Small Amplicons Mutation Library for Vaccine Screening by Error-Prone Polymerase Chain Reaction  

作　　者：程曼曼 张云龙 陈婷 张敏敏 陆昌瑞 CHENG Manman;ZHANG Yunlong;CHEN Ting;ZHANG Minmin;LU Changrui(College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China)

机构地区：[1]College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China

出　　处：《Journal of Donghua University(English Edition)》2023年第2期171-176,共6页东华大学学报（英文版）

基　　金：Shanghai Science and Technology Commission’s“Belt and Road Initiative”International Cooperation Project,China(No.19410741800)。

摘　　要：Library construction is a common method used to screen target genes in molecular biology.Most library constructions are not suitable for a small DNA library(<100 base pair(bp))and low RNA library output.To maximize the library’s complexity,error-prone polymerase chain reaction(PCR)was used to increase the base mutation rate.After introducing the DNA fragments into the competent cell,the library complexity could reach 109.Library mutation rate increased exponentially with the dilution and amplification of error-prone PCR.The error-prone PCR conditions were optimized including deoxyribonucleotide triphosphate(dNTP)concentration,Mn^(2+)concentration,Mg^(2+)concentration,PCR cycle number,and primer length.Then,a RNA library with high complexity can be obtained by in vitro transcription to meet most molecular biological screening requirements,and can also be used for mRNA vaccine screening.

关 键 词：error-prone polymerase chain reaction in vitro transcription DNA library RNA library 

分 类 号：Q523.8[生物学—生物化学]