福建农林大学学报（自然科学版）2017，Vol.46Issue(1)：81-88,8.DOI:10.13323/j.cnki.j.fafu(nat.sci.).2017.01.013
鲤鱼g型溶菌酶基因的cDNA克隆及其在毕赤酵母中的表达
cDNA cloning of g-type lysozyme gene from common carp and its expression in Pichia pastoris
摘要
Abstract
G-type lysozyme plays an important role in fish's immune response against microbial invasion. In the present study, a new g-type lysozyme isoform gene (CLYG) was separated from common carp (Cyprinus carpio) gill. The full-length cDNA of CLYG is 558 bp, which encodes 185 amino acid residues. The amino acid sequence deduced from the cDNA of CLYG contains 3 catalytic res-idues (Glu73, Pro86 and Asp97) without any signal peptide, which is similar to those of other teleost species. Subsequently, XhoⅠ and XbaⅠ restriction sites were added to the 5′ and 3′ ends of CLYG gene by PCR, respectively. After that, the expected fragment was ligated with expression vector pPICZαA, and followed by being transformed into competent Pichia pastoris X-33 cells. Then recombinant CLYG was induced in 1.0% methanol under pH 6.0 at 29℃ for 72 h. SDS-PAGE analysis and Western blot anal-ysis demonstrated that the recombinant CLYG was successfully expressed.关键词
鲤鱼/g型溶菌酶/毕赤酵母/重组表达Key words
common carp/g-type lysozyme/Pichia pastoris/recombinant expression分类
生物科学引用本文复制引用
李雯,陶妍..鲤鱼g型溶菌酶基因的cDNA克隆及其在毕赤酵母中的表达[J].福建农林大学学报（自然科学版）,2017,46(1):81-88,8.基金项目
上海市教育委员会产学研项目(15CXY30) （15CXY30）
农业部都市农业(南方)重点实验室开放项目(UA201307) （南方）
上海市交叉学科研究生拔尖创新人才培养平台项目(B5201120040) （B5201120040）
上海市2015高校内涵建设项目(A2018150009). （A2018150009）
