期刊论文

High Level Expression of Pectate Lyase Gene from Microbiology for Ramie Degumming and Its Bioinformatics Analysis

中文

华北农学报

1000-7091

2021

36

5

18-23

10.7668/hbnxb.20192340

1610242021002:中央级公益性科研院所基本科研业务费专项 2019JJ40332:湖南省自然科学基金 2019JJ50711:湖南省自然科学基金 ASTIP-IBFC08:创新工程项目 CARS-16-E22:现代农业产业技术体系建设专项 2019NK2071:湖南省重点领域研发计划 kq1901112:长沙市科技计划

本校为第一机构

为使苎麻生物脱胶果胶裂解酶PelG403高效表达。将pelG403从重组质粒pEASY-Blunt E1-G403更换至质粒拷贝数更高的pET28a载体中,导入Escherichia coli BL21(DE3)后进行诱导表达。采用DNS法进行酶活力测定,通过SDS-PAGE和Western Blot分析检验PelG403表达效果,并对其序列进行分析鉴定。结果表明: pET28a-G403 /BL21的果胶裂解酶活力为335.8 U/mL,较pEASY-Blunt E1-G403 /BL21提高了3.05倍;工程菌pET28a-G403 /BL21表达产物分子量比预测带His标签的融合蛋白分子量(43.6 ku)略小;PelG403含387个氨基酸,N末端前35个氨基酸为信号肽;理论pI值为7.64,有4个半胱氨酸,不稳定系数为27.42;其蛋白结构域含有典...

In order to efficiently express the pectate lyase PelG403 from ramie biodegumming. This study replaced pelG403 from the recombinant plasmid pEASY-Blunt E1-G403 to the pET28a vector with a higher plasmid copy number,and then introduced Escherichia coli BL21(DE3)to induce expression. The enzyme activity was determined by DNS method,and the expression effect of PelG403 was tested by SDS-PAGE and Western Blot analysis, and the sequence was analyzed and identified. The results showed that the pectate lyase activity of pET28a-G403 /BL21 was 335.8 U/m...