By continuing to use this site, you agree to the use of cookies. Mechanism of Cre-lox and FLP-FRT recombination Binding Synapsis , cleavage and strand exchange Three type of arrangement Inversion Translocation/ Insersion Deletion Application of Cre-lox and FLP-FRT recombination Disadvantage of FLP-FRT Advantage and disadvantage of Cre-lox Conclusion References KAUSHAL SAHU Follow Advertisement Advertisement Gene activation via Cre/lox-mediated excision in cowpea (Vigna systemhas two components: Cre recombinase derived from bacteriophage P1 two34- bp lox Crerecognizes; site-specificrecom- bination Cre-mediatedcatalysis re-ciprocal recombination between twolox bothtissue culture cells mice[18,19 . Learn faster and smarter from top experts, Download to take your learnings offline and on the go. Cre-Lox Recombination - CRI-LFDV Internship Promo Li A high-throughput screen identifying sequence and promiscuity characteristics of the loxP spacer region in Cre-mediated recombination. The Cre-lox system is used to introduce deletions, insertions, translocations, and inversions at a specific genomic locus. As an example, see 008917. These targeted DNA changes are useful in cell lineage tracing and when mutants are lethal if expressed globally. Well, it enables them to generate tissue-specific and inducible knockouts and thereby have exquisite control over the location and timing of gene expression important stuff when deleting a certain gene everywhere or during development leads to an embryonic lethal phenotype. orientation of the two loxP, one can create a gene inversion, translocation, or deletion. The loxP mutants are created to avoid these problems.[23]. eCollection 2022. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X03R618CYFR through backcrossing . Do not sell or share my personal information, 1. PubMed, Gu, H., Zou, Y.R., and Rajewsky, K. 1993. [29] This is done through the fusion of a mutated ligand binding domain of the estrogen receptor to the Cre recombinase, resulting in Cre becoming specifically activated by tamoxifen. The Cre-loxP recombination system is routinely used for the generation of mouse knockouts. Brault V, Besson V, Magnol L, Duchon A, Hrault Y. Handb Exp Pharmacol. Learn about the latest plasmid technologies and research tools. Why? In the lysogenic cycle the phage genome replicates with the rest of the bacterial genome and is transmitted to daughter cells at each subsequent cell division. For a more thorough introduction, check out Addgenes Cre-lox blog post. [28] Failure to activate both recombination events simultaneously confounds the interpretation of cell fate mapping results. [38], The P1 phage DNA when released into the host from the virion is in the form of a linear double stranded DNA molecule. PDF A Novel Cre/lox-Based Genetic Tool for Repeated, Targeted and cre-lox and cre recombinases in Mouse Genome Informatics (MGI): Module 2. Have you used the Cre-lox system in your research? A comparative analysis of microglial inducible Cre lines. See, for example, Lyznik et al., Site-Specific Recombination for Genetic Engineering in Plants, Plant Cell Rep, 21:925-932 (2003) and WO 99/25821, which are hereby incorporated by reference. In our study, we mutagenized 13 bp ( 5- ATAACTTCGTATA-3) of the right RBE of the loxP sequence. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. This review provides an overview of the cre-loxP system and its various permutations. National Library of Medicine Cre recombinase proteins bind to the first and last 13 bp regions of a lox site forming a dimer. The virus, called a prophage in such a state, accomplishes this via integration and excision. . Depending on the A novel maize variety designated X00R808 and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Here is the most efficient breeding scheme for generating Cre/loxtissue-specific or inducible knockouts. A novel maize variety designated 2ZZSB1121 and seed, plants and plant parts thereof are provided. OMP targeting vector with Cre FNF (neo-selectable marker flanked by FRT sites), M71 targeting vector with IRES CreFNF (neo-selectable marker flanked by FRT sites), Cre, Puro resistance and miRNA expression. BY KALYANI RAJALINGHAM Cre/lox regulated conditional rescue and inactivation with zebrafish . This tool is suitable for deleting antibiotic resistance genes, but above all it allows conditional knockouts that can be induced at specific times in the cell type of choice. And, it can be used to turn transgene expression on or off, track individual cells or cell lineages (as in theBrainboworConfettimice), generateinversionsortranslocations, and report gene expression. I highly recommend that you check out his blog. In tissues with no cre gene the target gene with be present and function normally. Here, we present a simple method that combines the use of CRISPR-Cas9 and Cre-LoxP for the Recombinant modified Cre recombinase protein translocates into the cytoplasm and nuc Additionally, Cre-containing adenovirus (Ad-Cre) or AAV (AAV-pgk-Cre) has been used to successfully introduce Cre into cells of interest. A conditional knockout (cKO) model is generated via several steps, using Cre-LoxP system as an example: The targeted fragment to be knocked out is flanked by LoxP elements. Read Addgene's Blog on Cre alternatives from Gerald Rubin's lab. (RBE); the two RBE are 13bp We've updated our privacy policy. I. Recombination between loxP sites. 2. The Cre/ lox system is one of the most powerful and versatile tools developed for mouse genetics. mutant/transgenic mice. A separate recombinase recognizes and binds to each of two recombination sites on two different DNA molecules or within the same DNA strand. We've encountered a problem, please try again. Chai W, Hao W, Liu J, Han Z, Chang S, Cheng L, Sun M, Yan G, Liu Z, Liu Y, Zhang G, Xing L, Chen H, Liu P. JBMR Plus. The limitations of creloxP technology and related considerations for experimental design are addressed, and alternative strategies for sitespecific genetic recombination and integration are discussed. Tap here to review the details. Engineered lox variants which differ on the spacer region tend to have varied but generally lower recombination efficiency compared to wildtype loxP, presumably through affecting the formation and resolution of recombination intermediate.[25]. Editing, Cloning When there are We've updated our privacy policy. Now customize the name of a clipboard to store your clips. Two plasmids can be joined using the variant lox sites 71 and 66. The following text is excerpted from Siegel et al. Some floxed alleles are more easily recombined than others. The Cre-lox system serves several functions in the phage: it circularizes the phage DNA into a plasmid, separates interlinked plasmid rings so they are passed to both daughter bacteria equally and may help maintain copy numbers through an alternative means of replication. CreER(T2) resides within the cytoplasm and can only translocate to the nucleus following tamoxifen administration, allowing tight temporal control of recombination. 8600 Rockville Pike The fluorescent reporter cassette will contain a promoter to permit high expression of the fluorescent transgene reporter (e.g. FEBS Lett. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. 4. How do I prepare and deposit my plasmids? US Patent for Maize hybrid X03R618CYFR Patent (Patent # 11,589,540 Among these are loxN (GtATACcT), lox2272 (GgATACtT), and lox511 (GtATACAT). Have questions about your order, deposit, or a plasmid? In this example, expression of Cre recombinase removes the transgene (Figure 6). Once tamoxifen is introduced, it is metabolized into 4-hydroxytamoxifen, which then binds to the ER and results in the translocation of the CreER into the nucleus, where it is then able to cleave the lox sites. he canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT. A soybean cultivar designated 03070412 is disclosed. Cre recombinase was used to mediate recom- Introduction bination between a chromosomally introduced loxP se- quence in Arabidopsis thaliana (35S-lox-cre) and trans- Site-specific recombination systems from prokaryotes ferred DNA (T-DNA) originating from Agrobacterium and yeast, such as the bacteriophage P1 Cre/lox system, tumefaciens (plox-npt), NNNTANNN", where "N" indicates which bases may vary from the canonical sequence, Sternberg, N. and Hamilton, D. 1981. The use of the Cre/loxP system for generating tissue specific or inducible knockouts is a powerful tool for mouse genetics, but it is only one piece of the total picture. A novel maize variety designated X03R618CYFR and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Approximately 25% of the progeny from this mating will be homozygous for theloxP-flanked allele and hemizygous/heterozygous for thecretransgene. Expressed in Muller glia in the retina. Activate your 30 day free trialto continue reading. For two lox sites on the same chromosome arm, inverted loxP sites will cause an inversion of the intervening DNA, while a direct repeat of loxP sites will cause a deletion event. Explaining these schemes can be difficult, and I swear that I can sometimes feel customers eyes cross while Im trying to explain them over the telephone. & ORFs. Jae Lee-Exam.docx - <The exam for Jae Lee's sessions> Approximately 50% of the offspring will be heterozygous for theloxPallele and hemizygous/heterozygous for thecretransgene. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. The Cre- lox recombination system is derived from the bacteriophage P1. Site-specific recombination is also an important process that viruses, such as bacteriophages, adopt to integrate their genetic material into the infected host. The present invention relates generally to transgenic plants. Plasmids. b) The second step is supply of Cre recombinase. Unauthorized use of these marks is strictly prohibited. Since removal of the stop cassette is permanent, the reporter genes are expressed in all the progeny produced by the initial cells where the Cre was once activated. Do you have any tips or tricks, or clever cre-dependent techniques? Cre activates gene of interest, removes Neo cassette. The asymmetry in the core sequence gives the loxP site directionality, and the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT. The FLP-FRT technology can be an effective alternative to Cre-lox, and has also been used in conjunction with it, allowing for two separate recombination events to be controlled in parallel. sites contain nucleotide variations in the left or right arms, respectively named LE and RE. Bacterial genome editing by coupling Cre- lox and CRISPR-Cas9 - PLOS See plasmid, Cre activates gene of interest by removing Stop sequences. Plasmids 101, Research mouse models that incorporate the Cre/loxsystem typically involve generating double,triple, quadruple, or evenn-tuple(is that really a word?) Cre-Lox recombination is a sophisticated site-specific recombinase technology that allows DNA modification to be targeted to a specific cell type or to be triggered by a specific external stimulus. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. 1. Conditional gene inactivation can be achieved with the Cre/ lox system in which the bacteriophage Cre recombinase promotes site-specific recombination at compatible loxP sites engineered in a gene of interest ( Sauer and Henderson, 1988 ). An additional level of control can be achieved by using his Cre recombinase, engineered to reversibly activate in the presence of the estrogen analogue 4-hydroxy tamoxifen. AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR, AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR, AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR, AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA, pGEMT-Pax7bait-P2a-GAP43-Cherry-T2a-mERt-Cre-Ert, pCAGGS-mTagBFP2-T2A-sfGFP-GSAx9-iCre-ERT2, pCAG-Synaptophysin-TdTomato-IRES2-OHT-Cre, hsp70l-loxP-mCherry-STOP-loxP-H2B-GFP_cryaa-cerulean, pJFRC172-10XUAS-loxP>-dSTOP-loxP>-myr::GFP, Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientists View, CREM and CMV-STOP-luc cassette, separated by insulator, Cre-EGFP fusion in a promoterless vector with several available restriction sites, tet inducible Cre-GFP fusion, metallothionein MT-I region including the polyadenylation site and several introns, Cre-ERT2 with loxp cassette; Targeting vector for Rosa26 locus, CREM fused to mutant AR ligand binding domain, Cre fused to HIV-TAT - promotes cellular uptake of recombinant Cre, Cre-Myc, Expressed in rod photorecetor cells in the retina. Inducible Cre activation is achieved using CreER (estrogen receptor) variant, which is only activated after delivery of tamoxifen. First, let us focus on the Cre-loxP system which requires a Cre In more recent years, however, this view has been challenged, and most of the current models for Int, Xer, and Flp recombination involve only limited branch migration (13 base pairs of the Holliday intermediate), coupled to an isomerisation event that is responsible for switching the strand cleavage specificity. 1992. Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells. Special features include: Use the search box below to find the perfect Cre expression construct for your experiment. Now, one point of crossover has occurred, which is what is called a Holliday Intermediate. the Flp-FRT system. Orban, P.C., Chui, D., and Marth, J.D. [7] Combining these advances in collaboration, the laboratories of Drs. Tetracycline-inducible Expression Systems New - Do you have suggestions for other plasmids that should be added to this list? The Cre/lox system can also be used to produce strains in which a transgene is either inducible or expressed only in certain tissues. Thus, the orientation, and location of the loxP sites are important determinants of FLP-FRT & Cre-lox Recombination This breeding scheme may not be the most efficient one for producing all Cre/loxstrains. These lox variants only proceed recombination with the same type of lox sites, with no ability to interact with the other types. Plasmid Elements, Expressed in chondrocytes, mCherry and Cre-ERT2 - Tamoxifen inducible; Targeting vector for Pax7, Venus and Cre-ERT2 with MCS for inserting promoter, Emerald and Cre-ERT2 with MCS for inserting promoter, TFP and Cre-ERT2 with MCS for inserting promoter, tdTomato and Cre-ERT2 with MCS for inserting promoter, Venus, Cre-ERT2, targeting vector with MCS for homology arms, Emerald, Cre-ERT2, targeting vector with MCS for homology arms, TFP, Cre-ERT2, targeting vector with MCS for homology arms, tdTomato, Cre-ERT2, targeting vector with MCS for homology arms, GAG-Cre fusion; for the production of VLPs loaded with CRE protein, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, unskippable linker) - Tamoxifen inducible, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, GSAx9 linker) - Tamoxifen inducible, iCre-ERT2 (PAPGSTMA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PA N-terminus) - Tamoxifen inducible, CreLite - Red light-inducible Cre; N terminus Cre fused to PIF6, CreLite - Red light-inducible Cre; C terminus Cre fused to PhyBdelta, CreLite system components, PhyBdeltaCreC and PIF6CreN, in middle entry vector (Tol2 kit), CreLite; Tol2 destination vector with mTagBFP2, CreLite system components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer vector, CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector, Cre recombinase split with Vivid photodimers, Cre-ERT2 - Tamoxifen inducible; AAV donor vector, iCre-ERT2 (PAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSAS N-terminus) - Tamoxifen inducible, N-terminal Cre component split with Vivid photodimers, C-terminal Cre component split with Vivid photodimers, Split Cre fused to nuclear localized wild-type VVD, Retinal ganglion cell-specific expression of Cre, Cre activates shRNA expression, removal of EGFP; See also similar plasmids, Cre turns off shRNA expression, removal of EGFP; See also similar plasmids, Cre activates gene of interest by removing Stop sequences; Puro selection. Cre can invert a region rather than delete it. place. two loxP sites in the same direction Activate your 30 day free trialto unlock unlimited reading. This provides the advantage that the Cre recombinase is active for a short time. In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements promoters, origins of replication, protein tags, and antibiotic resistance markers(just to name a few). cre-lox and cre recombinases in Mouse Genome Informatics (MGI): Module 2 MouseGenomeInformatics 17.7k views Knock out technology (final) Dr Vijayata choudhary 22.2k views Gene knockout GGS Medical College/Baba Farid Univ.of Health Sciences. When cells that have loxP sites in their genome express Cre, a recombination event can occur between the loxP sites. 2. Principle of Cre-DIO system - GeneMedi Phages like the lambda phage use their site specific recombinases to integrate their DNA into the host genome during lysogeny. In this fashion, opposite DNA strands are joined together. Cre-loxP recombination that result in insertion requires loxP sites to flox the gene of interest, with the loxP sites oriented towards each other. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. This has been largely due to the simplicity of its use and its adaptability to address diverse biological questions. A system of site-specific recombination (q.v.) FEBS Lett. The protein Cre recombinase recognizes 34 bp loxP sites, and the orientation and location of the loxP sites determines how the genetic material will be rearranged. Cre/loxP Recombination System: Applications, Best Practices - Taconic If the two loxP sites are in the same orientation, the floxed sequence (sequence flanked by two loxP sites) is excised; however if the two loxP sites are in the opposite orientation, the floxed sequence is inverted. 2001 Jun 15;499(1-2):147-53. doi: 10.1016/s0014-5793(01)02541-8. Cre-loxP - Wikipedia (Already know this? a) The chromosomal copy of the target gene replaces the target gene flanked by loxP sites. The Saccharomyces cerevisiae FLP-FRT recombination system is another site-directed recombination technology very conceptually similar to Cre-lox, with flippase (Flp) and the short flippase recognition target (FRT) site being analogous to Cre and loxP, respectively. LBD. The Cre enzyme targets loxP sites at the ends of this molecule and cyclises the genome. Missirlis PI, Smailus DE, Holt RA. recombination is reversible, strategies for irreversible integration rely on mutated . At the given specific site on the DNA, the hydroxyl group of the tyrosine in the recombinase attacks a phosphate group in the DNA backbone using a direct transesterification mechanism. [2], Cre-Lox recombination is a special type of site-specific recombination developed by Dr. Brian Sauer and patented by DuPont that operated in both mitotic and non-mitotic cells, and was initially used in activating gene expression in mammalian cell lines. Embodiments include the seeds of soybean 4826008, the plants of soybean 4826008, to plant parts of soybean 4826008, and methods for producing a soybean plant produced by crossing soybean 4826008 with itself or with another soybean variety. By whitelisting SlideShare on your ad-blocker, you are supporting our community of content creators. More specifically, it relates to methods and compositions for the introduction of DNA using circular molecules that are not able to replicate outside a host cell. Activate your 30 day free trialto continue reading. To rule out the possibility that the gene recombination efficacy was insufficient for CK19;PTEN / mice to develop tumors, we administered the same dose of TAM for 3 consecutive days by oral gavage, but no ampullary tumors were observed after 90 days (n = 4). The Cre-lox system is a technology that can be used to induce site-specific recombination events. cre lox recombination ppt Common construct types include: Note: Through a partnership with genOway, Addgene is able to distribute materials containing FLEx technology. (<U>G</U>ene-<U>A</U>uto-<U>E</U>xcision <U>B</U>inary <U>S</U>ystemGAEBS)(transcription system) . Cre-lox recombination, discovered in the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox based technologies are still being developed today. Explanation: The Cre recombinase protein of bacteriophage P1 mediates site-specific recombination at a 34 bp sequence, loxP. Protocol for Cre Recombinase (M0298) | NEB LoxP (locus of X(cross)-over in P1) sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats separated by an 8-bp long asymmetric core spacer sequence. The Cre recombinase is an enzyme that is required for KAUSHAL KUMAR SAHU Cre- loxP cre loxP GFP cre GFP cre cre cre Now researchers are using sequencing technology to better, Assessing patient health risks with limited or unknown family history, Using the power of sequencing on our smallest organisms. Read the genOway press release for more information. Disclaimer. Transgenic mice containing a gene surrounded by loxP sites are mated with transgenic mice that have the cre gene expressing only in one cell type. How can I be notified when a plasmid from a specific lab or paper is available? The C domain is similar in structure to the domain in the Integrase family of enzymes isolated from lambda phage. Heteromeric . For example, mating the transgenic strain in Figure 1 to a strain that expresses Cre recombinase in mammary tissue produces double transgenic offspring that express theKras oncogene only in the mammary glands (Figure 5). Maize hybrid X13C733 is an invention by Mario Rosario Carlone, Jr., Princeton IL UNITED STATES. The two loxP sites then align in a parallel orientation, allowing the four Cre proteins to form a tetramer. This reaction is aided with the endonuclease activity of an enzyme. Cre (shown at left) is a bacteriophage P1 member of the integrase family, catalyzing site-specific recombination between two, 34-base pair lox DNA sequences. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. ADVERTISEMENTS: lox. If you are looking for the right Cre-expressing strain for your experiments, you can search for strains at the JAX Cre Repository or through the Cre Portal at MGI. Cre, or cyclic recombinase, is one such enzyme. 2007;(178):29-48. doi: 10.1007/978-3-540-35109-2_2. US Patent for Maize inbred 2ZZSB1121 Patent (Patent # 11,589,535 issued Recombination/Bacteriophage P1-derived Cre-lox - parts.igem.org Cre-lox, . All cell types tested in mice have been shown to undergo transgenic Cre recombination. An attP site on the phage exchanges segments with an attB site on the bacterial DNA. Cre-lox in vivo/vitro . You may not be able to create an account or request plasmids through this website until you upgrade your browser. PubMedPMID:6276558. Cre Recombinase - 1 ul (1 unit*) H20 - Xul (up to 50ul) Incubate at 37C for 30 minutes and then 70C for 10 minutes. Translocation: If the sites are on separate DNA molecules, a translocation event is generated at the loxP sites. The P1 phage is a temperate phage that causes either a lysogenic or lytic cycle when it infects a bacterium. 2023 Feb 10;12(4):568. doi: 10.3390/cells12040568. Choose the incorrect statement for cre-lox excision. The system utilizes Cre recombinase, a bacteriophage P1 enzyme that recognizes a short target DNA sequence ( loxP site). Cre-LoxP Recombination Hannah Park 316 subscribers Subscribe 2.9K 317K views 7 years ago This video discusses the mechanisms behind the Cre-LoxP Recombination, which has been used in many. To verify that Cre recombinase expression does not contribute to a phenotype of interest, I suggest using thecrestrain itself (without anyloxP-flanked alleles) as a control (Figure 3B). This can also take place in the absence of the Cre lox system[39] with the help of other bacterial and viral proteins. Fill out our Suggest a Plasmid form or e-mail [emailprotected] to help us improve this resource! The Cre recombinase is fused to a mutant form of the oestrogen receptor, which binds the synthetic oestrogen 4-hydroxytamoxifen instead of its natural ligand 17-estradiol. Cre is a 38 kDa recombinase protein from bacteriophage P1 that catalyzes recombination between LoxP sites. Mol Urol. PubMed, Orban, P.C., Chui, D., and Marth, J.D. Matsumura T, Watanabe T, Totsugawa T, et al. Cre-loxP-Mediated Recombination: General Principles and - PubMed Maize hybrid X13C733 Patent | Application Number: 13396674 This system has allowed researchers to manipulate a variety of genetically modified organisms to control gene expression, delete undesired DNA sequences and modify chromosome architecture. Click here to review the details. Targeting Your Dna With the Cre/Lox System | Scq [13][14] To date, NIH Blueprint for Neuroscience Research Cre projects have created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. By HHS Vulnerability Disclosure, Help recombination; recombination between two loxP sites can induce a deletion. We archive and distribute high quality plasmids from your colleagues. The Cre protein (encoded by the locus originally named as "Causes recombination", with "Cyclization recombinase" being found in some references)[21][22] consists of 4 subunits and two domains: The larger carboxyl (C-terminal) domain, and smaller amino (N-terminal) domain.

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