[Isothermal unidirectional elongation method of gene synthesis].
Gene synthesis is gaining popularity as a versatile means of cloning genes. The prevailing synthesis methods are PCR-based and ligase-based gene synthesis. Both of them are not independent of the sequence content of the target gene. In the present study, a new gene synthesis method-isothermal unidirectional elongation method (IUEM) is proposed, which involves the cooperation of three enzymes, and is independent or partially independent of the target gene sequence by the specially designed oligonucleotides that form hairpins. The synthesis process is isothermal and serial in IUEM. Several factors that affect the elongation were studied, and two DNA segments of length 254 bp and 300 bp were synthesized with IUEM. The synthesized segments could work as building blocks for the synthesis of longer DNA sequences.
Lin
Ji-Wei
JW
Bio-X Life Science Research Center, Life Science and Technology College, Shanghai JiaoTong University, Shanghai 200030, China. ljwopen@gmail.com
Zhang
Xiao-Dong
XD
Cao
Xue-Yan
XY
Hu
Jun
J
Ji
Cong-Liang
CL
D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D005813:Genes, Synthetic / Q000235:genetics*; D008954:Models, Biological; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013696:Temperature
D004247:DNA