食品科学2017，Vol.38Issue(10)：31-36,6.DOI:10.7506/spkx1002-6630-201710006
枯草芽孢杆菌纤维素酶基因整合载体的构建
Construction of Cellulose Gene Integration Vector of Bacillus subtilis
摘要
Abstract
Aim:To obtain an engineered bacterial strain able to express cellulose-degrading enzymes through constructing an integration vector using Bacillus subtilis as the host.Methods:The homologous fragments M1 and M2 were cloned by polymerase chain reaction (PCR) from the genome of B.subtilis LN.The glucosidase gene CelKg,homologous fragments M1 and M2 and the strong promoter P43 were ligated to the pGEM-T vector by T4 DNA Ligase to construct the integrated vector pGEM-Kmpgrnt.The vector was then transformed to B.subtilis LN by double crossover homologous recombination method.Results:The integration vector was successfully constructed and integrated into the genome of B.subtilis LN,as verified by PCR.Congo red staining indicated that the recombinant B.subtilis could obviously degrade sodium carboxymethyl cellulose in the medium.The cellulase activity from the culture supematant harvested after 18 h culture of the recombinant strain in a modified medium at 37 ℃ with shaking was increased by 115％ as compared with the wild-type B.subtilis LN.关键词
同源重组/纤维素酶基因/枯草芽孢杆菌Key words
homologous recombination/cellulase/Bacillus subtilis分类
生物科学引用本文复制引用
聂利波,王占彬,史敦胜,宋洋洋,李旺..枯草芽孢杆菌纤维素酶基因整合载体的构建[J].食品科学,2017,38(10):31-36,6.基金项目
国家自然科学基金青年科学基金项目(31101744) （31101744）
河南省重大科技专项(131100110300) （131100110300）
