[Synthesis of total sFat-1 gene by PCR-restrict enzyme ligation method].
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.
Zhu
Gui-Ming
GM
College of Animal Science and Technology, Northwest A&F University, Yangling, China.
Chen
Hong
H
Lu
Jian-Shen
JS
Zhou
Yan-Rong
YR
Wu
Xiao-Jie
XJ
Chen
Hong-Xing
HX
Deng
Ji-Xian
JX
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
D012313:RNA; D044943:Fatty Acid Desaturases; C078565:omega-3 fatty acid desaturase; D004262:DNA Restriction Enzymes