摘要
根据人ＩＣＦ－１（Ｈｕｍａｎ Ｉｎｓｕｌｉｎ－ｌｉｋｅ Ｇｒｏｗｔｈ Ｆａｃｔｏｒ－１，ｈＩＧＦ－１）的天然基因序列，在尽量保存原基因序列的基础上，将一些密码子换成Ｐｉｃｈｉａ ｐａｓｔｏｒｉｓ酵母的偏爱密码子，人工合成ｈＩＧＦ－１双链ＤＮＡ．将此合成基因整合人Ｘ－３３酵母的染色体上，用Ｚｅｏｃｉｎ＋平板筛选得到重组菌株．以胞外分泌的方式表达了ｈＩＧＦ－１蛋白，表达产物经聚丙烯酰胺凝胶电泳和ｗｅｓｔｅｒｎ－ｂｌｏｔ分析，得到该表达产物的相对分子质量约为７２００．
According to the known sequence of hICF-1 cDNA, we synthesized the double-chain DNA of hIGF-1 by changing the natural code of hIGF-l amino to the favorite code of Pichia pastoris on the base of preserving its natural DNA sequence as closely as possible. We transformed the yeast cell-X-33 with this synthesized gene and this gene was integrated into the chromosome of the yeast. The straira of the recombinant yeast were screened with the Zeocin+ plate. The product was secreting expressed and its molecular weight was determined as 7200 by SDS-PACE and Western-blot. The amount of unpuri- fied protein was about 49% of the total protein in the supernatant of the culture media.
出处
《中山大学学报（自然科学版）》
CAS
CSCD
北大核心
2000年第z2期44-47,共4页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
国家1035资助项目
作者简介
彭鸿娟(1971～)，女，讲师.
