降解乳蛋白菌株中蛋白酶基因的克隆及分析
China Dairy Industry(2010)
Abstract
从降解乳蛋白菌株Bacillus subtilis strain ZJ06中克隆蛋白酶基因apr,分析蛋白酶基因apr结构,预测蛋白酶性质。以PCR方法从降解乳蛋白菌株Bacillus subtilis strain ZJ06基因组中扩增一碱性蛋白酶基因apr片段,对其克隆测序;提取菌体总RNA,进行RT-PCR;地高辛随机引物法标记探针,进行菌落原位杂交。结果表明,从降解乳蛋白菌株Bacillus subtilis strain ZJ06基因组中克隆到碱性蛋白酶基因apr,ORF1146bp编码382个氨基酸,理论分子量为39 ku,信号肽长30个氨基酸,是分泌型蛋白酶,蛋白酶原352个氨基酸,成熟酶275个氨基酸,GC含量51.4%,理论分子量为27.4 ku。RT-PCR、菌落原位杂交方法证实该基因的存在与表达。
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Key words
clone,alkaline protease gene,analyses.,milk-protein degradation
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