Template-directed chemical ligation to obtain 3'-3' and 5'-5' phosphodiester DNA linkages.
Up to now, the direct ligation of two DNA fragments with opposite directions to obtain 3'-3' or 5'-5' phosphate ester bonds is still challenging. The only way to obtain DNA oligonucleotides containing a 3'-3' or 5'-5' inversion of polarity sites is based on professional DNA chemical synthesis. Herein, we demonstrate a convenient template-directed chemical ligation that enables 3'-3' and 5'-5' linkages of two DNA oligonucleotides. This method is based on the assembly of two oligonucleotides on a template in opposite directions through forming antiparallel and parallel duplexes simultaneously, followed by coupling with N-Cyanoimidazole under mild condition. Moreover, on the basis of DNA oligonucleotides with 5'-5' linkage obtained through our template-directed chemical ligation, we developed a new cDNA display technique for in vitro selection of functional polypeptides.
1] Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China [2] The College of Life Science, Sichuan University, Chengdu, Sichuan, 610065, China.
Haodong
H
Chen
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China.
Feng
F
Du
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China.
Gangyi
G
Chen
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China.
Frank
F
Streckenbach
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China.
Afshan
A
Yasmeen
The College of Life Science, Sichuan University, Chengdu, Sichuan, 610065, China.
Yun
Y
Zhao
1] Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan, 610041, China [2] The College of Life Science, Sichuan University, Chengdu, Sichuan, 610065, China.
Zhuo
Z
Tang
D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism; D007093:Imidazoles / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*
D007093:Imidazoles; D009841:Oligonucleotides; D011088:DNA Ligases; D000072481:DNA Ligase ATP; C051673:N-cyanoimidazole