A novel high-throughput (HTP) cloning strategy for site-directed designed chimeragenesis and mutation using the Gateway cloning system
نویسندگان
چکیده
منابع مشابه
A novel high-throughput (HTP) cloning strategy for site-directed designed chimeragenesis and mutation using the Gateway cloning system
There is an increasing demand for easy, high-throughput (HTP) methods for protein engineering to support advances in the development of structural biology, bioinformatics and drug design. Here, we describe an N- and C-terminal cloning method utilizing Gateway cloning technology that we have adopted for chimeric and mutant genes production as well as domain shuffling. This method involves only t...
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SFL1 and SFl2 (SETH Four Like) genes are two members of SETH4 gene family in Arabidopsis thaliana expressed in saprophytic tissues. In this study, expression of SFL1 and SFL2 genes were studied using Gateway Cloning Technology. Primers were designed for PCR amplification of promoter region of SFL1 (900 bp) and SFL2 (930 bp) genes having attB1 recombination sites using Kod Hi Fi DNA polymerase e...
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Direct cloning of PCR fragments by TA cloning or blunt end ligation are two simple methods which would greatly benefit high-throughput (HTP) cloning constructions if the efficiency can be improved. In this study, we have developed a ribosomal binding site (RBS) switching strategy for direct cloning of PCR fragments. RBS is an A/G rich region upstream of the translational start codon and is esse...
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The current challenge, now that two plant genomes have been sequenced, is to assign a function to the increasing number of predicted genes. In Arabidopsis, approximately 55% of genes can be assigned a putative function, however, less than 8% of these have been assigned a function by direct experimental evidence. To identify these functions, many genes will have to undergo comprehensive analyses...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 2005
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/gni103