title: [Synthesis of total sFat-1 gene by PCR-restrict enzyme ligation method].
abstract: Gene synthesis is very important in life science research, and it becomes a technique in common use. It is difficult for long gene synthesis, because the mismatches and mutations of DNA sequence in nucleotide fragments assembling. This study established a new method for long gene synthesis, which was referred to as PCR-restrict enzyme ligation method. With this method, a omega-3 fatty acid desaturase gene, sFat-1, from Caenorhabditis briggssae, was successfully assembled from 27 synthesized nucleotide fragments (60 ~ 68 bp for each fragment ) following 3 rounds of PCR (7 reactions) and 2 rounds of restrict enzyme ligation (3 reactions). This shows that the PCR-restrict enzyme ligation method is an effective method for long gene synthesis.
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, China.
authors.forename: Gui-Ming
authors.initials: GM
authors.lastname: Zhu
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Jian-Shen
authors.initials: JS
authors.lastname: Lu
authors.forename: Yan-Rong
authors.initials: YR
authors.lastname: Zhou
authors.forename: Xiao-Jie
authors.initials: XJ
authors.lastname: Wu
authors.forename: Hong-Xing
authors.initials: HX
authors.lastname: Chen
authors.forename: Ji-Xian
authors.initials: JX
authors.lastname: Deng
mesh_terms: D000818:Animals; D001483:Base Sequence; D002107:Caenorhabditis / Q000235:genetics*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism*; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000235:genetics
chemical_list: D012313:RNA; D044943:Fatty Acid Desaturases; C078565:omega-3 fatty acid desaturase; D004262:DNA Restriction Enzymes