%0 Journal Article
%T Amplification and bacterium expression of high-molecular-weight glutenin subunit 1Bx14 gene from wheat<br>小麦高分子量谷蛋白14亚基基因的PCR体系及原核表达研究
%A JIN Wei-bo
%A WU Fang-li
%A GUO Ai-guang
%A <br>金伟波
%A 吴方丽
%A 郭蔼光
%J 浙江大学学报(农业与生命科学版)
%D 2007
%I Zhejiang University Press
%X HMW-GS 1Bx14 was candidated as a good subunit for dough quality. HMW-GS 1Bx14 coding sequence can be amplified and expressed in vitro for characterizing its function. An improved PCR method was presented for amplification of HMW-GS 1Bx14 which is a fragment of large (about 2.4 kb) and contain GC-rich regions. This experiment adapted following three stratages to optimize PCR system for successful amplification: i)designing a couple of primers with high Tm; ii)Adopting two-temperature PCR; iii) Improving PCR buffer for beneficial long amplicon; adding some organic solvents(DMSO, Lycine) and BSA for resistant to form secondary structure and lengthen in polymerase half-life, respectively. Furthermore, it is very difficulty to express HMW-GS 1Bx14 in E.coli because of the bias of codon usage. The Rosetta(DE3)plysS bacterium was used to conquer the effection of rare codon and express HMW-GS 1Bx14 gene successfully. It will be benefit for further studying on the function of 1Bx14 gene.
%K wheat
%K HMW-GS 1Bx14
%K PCR system
%K bacterium expression<br>小麦
%K 14亚基
%K PCR体系
%K 原核表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=03F54A49DE00578AA0E5DDF5BC021AA7&cid=298920A27C9BAA22346FCA384240FAA4&jid=466C05D88593309B3EB5CF34361E6426&aid=5FDC770A04082CA5&yid=A732AF04DDA03BB3&vid=27746BCEEE58E9DC&iid=CA4FD0336C81A37A&sid=94E7F66E6C42FA23&eid=771152D1ADC1C0EB&journal_id=1008-9209&journal_name=浙江大学学报(农业与生命科学版)&referenced_num=1&reference_num=7