基质金属蛋白酶-2(明胶酶A)在肿瘤侵袭和转移中具有重要作用,为进一步研究其作用机制,在酵母系统中表达了金属蛋白酶-2通过PCR方法获得明胶酶A的表达序列,酶切和测序结果证明序列正确后,构建明胶酶A的表达载体pPIC9/GelA,电击法转化酵母得到阳性克隆.明胶酶谱分析说明表达产物具有明胶底物特异性,SDS-聚丙烯酰胺凝胶电泳表明,表达产物的分子质量为66 ku.
Matrix metalloproteinase-2 (MMP-2) plays important role in tumor invasion and metastasis. MMP-2 is expressed in yeast expression system for further studying its function mechanism. The expression sequence of MMP-2(gelatinase A) have been obtained by PCR amplifying,restriction enzyme cut and sequencing analysis demostrate that the sequence is correct. The expression vector pPIC9/GelA containing gelatinase A sequence is constructed and transformed the yeast Pichia pastoris. The gelatin enzymography demostrate that recombinant protein secreted by transformant yeast is capable of degrading gelatin and the molecular weight of the matrix metalloproteinase-2 is 66 ku in SDS-PAGE.
丁秀云,李春海,朱运峰,孙丽亚,颜春洪,张立新,冯东晓,陈高明.基质金属蛋白酶-2在酵母系统中表达的初步研究[J].生物化学与生物物理进展,1999,26(3):291-294
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