抗菌肽天蚕素B基因及其串联体在毕赤酵母中的表达  

Antibacterial peptide Cecropin B and its tandem gene expressed in Pichia pastoris

作　　者：王秀青[1] 张素芳[2] 曹瑞兵[1] 周斌[1] 陈溥言[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095 [2]中国科学院大连化学物理研究所,辽宁大连116023

出　　处：《南京农业大学学报》2007年第3期120-123,共4页Journal of Nanjing Agricultural University

基　　金：江苏省科技攻关项目(BE2006364);教育部高等学校博士点基金(20050307023)

摘　　要：通过重叠区扩增基因拼接法(SOE)合成抗菌肽天蚕素B基因,并在其N端引入Kex2酶切位点。亚克隆天蚕素B并将3个亚克隆串联在一起,每个单体前都加上Kex2酶切位点,将天蚕素B以及串联体克隆至表达载体pPICZαA上,用SacⅠ酶切使之线性化,采用电击法转化毕赤酵母SMD1168,转化子用小瓶发酵。经Tricine-SDS-PAGE检测,在α信号因子的引导下,表达产物可以分泌到培养基中,且具有明显抑菌活性。Cecropin B gene was achieved through the gene splicing by overlap extension （SOE）. Especially a Kex2 signal cleavage site was fused in N end of the antibacterial peptide gene. Cecropin B gene was subcloned and ligated tandem. The modified Cecropin B and its tandem genes cloned into the pPICZαA vector to construct the recombinant expression vectors. The recombinant expression vectors were linearized by Sac I and transformed into Pichia pastoris SMD1168 strain by electroporation. The positive clones were screened and those clones were fermented in flask. Tricine-SDS-PAGE showed that the Cecropin B protein could be se- creted into the culture leading by α-factor from pPICZcαA. Antibacterial activities and heat-stability were also found.

关 键 词：天蚕素B 抗菌肽 毕赤酵母 表达 抑菌活性 重叠区扩增基因拼接法(SOE) 

分 类 号：Q784[生物学—分子生物学]