摘要
将抗真菌蛋白Ｒｓ－ＡＦＰ１和Ｒｓ－ＡＦＰ２全长ｃＤＮＡ插入表达质粒ｐＥＴ－２２ｂ／ＮｃｏＩ＋ＳａｃＩ位点，构建成融合蛋白表达载体ｐＲＡＦ１和ｐＲＡＦ２．将不含信号肽编码序列的Ｒｓ－ＡＦＰ１和Ｒｓ－ＡＦＰ２ｃＤＮＡ分别插入ｐＥＴ－２２ｂ／Ｎｃｏｌ＋Ｓａｃｌ和ｐＥＴ－２２ｂ／Ｎｄｅｌ＋ＳａｃＩ位点，构建成不含信号肽序列的融合蛋白表达载体ｐＲＡＦ３、ｐＲＡＦ４和非融合蛋白表达载体ｐＲＡＦ５和ｐＲＡＦ６．将构建的上述各种表达载体转化Ｅ．ｃｏｌｉＢＬ２１，挑菌落培养，ＩＰＴＧ诱导，使Ｒｓ－ＡＦＰｓ基因得到表达，并用体外抑菌试验检测表达产物的活性，结果表明，各种表达载体的表达产物均具有不同程度的抑菌活性，其中，ｐＲＡＦ３和ｐＲＡＦ４表达产物的抑菌活性较明显．
Abstract Three groups of expression vectors were constructed in this study ,which were ① pRAF 1 and pRAF 2,the expression vectors of fusion protein encoded by part sequence of signal peptide of pET 22b and the full length Rs AFP 1 or Rs AFP 2 cDNA; ② pRAF 3 and pRAF 4, the expression vectors of fusion protein encoded by part sequence of signal peptide of pET 22b and the sequence of mature peptide of Rs AFP 1 or Rs AFP 2;③ pRAF 5 and pRAF 6,the expression vectors of non fusion protein encoded only by the sequence of mature peptide of Rs AFP 1 or Rs AFP 2. Rs AFPs genes were expressed in higher level after the above mentioned expression vectors transferred into E.coli BL21 and induced by IPTG.The expression products showed biological activity in considerable levels as well.
出处
《生命科学研究》
CAS
CSCD
1998年第1期17-23,共7页
Life Science Research
基金
农业部"九五"生物技术重点项目
