一种新的等温单向生长基因合成法初步探索.pdfVIP

HEREDITAS (Beijing) 2007 6 , 29(6):765770 ISSN 0253-9772 DOI: 10.1360/yc-007-0765 1 1 1 1, 2 , , , 1., Bio-X , 200030; 2. , 201800 : PCR P (L ) — (Isothermal Unidirectional Elongation Method, IUEM), 3 , DNA , , , , 254 bp 300 bp DNA : ; ; ; Isothermal unidirectional elongation method of gene synthesis 1 1 1 1,2 LIN Ji-Wei , ZHANG Xiao-Dong , CAO Xue-Yan , HU Jun 1. Bio-X Life Science Research Center, Life Science and Technology College, Shanghai JiaoTong University, Shanghai 200030, China; 2. Shanghai Institute of Applied Physics, Chinese Academy of Sciences, P. O. Box 800-204, Shanghai 201800, China Abstract: 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. Keywords: gene synthesis; isothermal; unidirectio