摘要
根据枯草杆菌(Bacillus subtilis)基因文库中的淀粉酶基因的碱基序列,设计引物。以枯草杆菌菌体DNA为模板,利用PCR扩增出分子量大约为2.3 kb的淀粉酶基因片段。借助于核酸内切酶和连接酶把该基因植入到pHMSXD1质粒中,构建pHMSXD2质粒载体。通过高压电击处理把载体植入到大肠杆菌(JM10)中并成功表达。该株大肠杆菌在酶基因表达后,其淀粉酶活力达到1.037 6 U/mL(LB培养液)和1.361 0 U/mL(矿物元素培养液),分别比原始的枯草杆菌的淀粉酶活力提高了46倍和286倍。
Amylase gene （2.3 kb） form Bacillus subtilis was amplified by polymerase chain reaction （PCR）. After the amylase genes were connected with a plasmid （pHMSXD1） and transformed into Escherichia coli （E.coli） by electroporation, the transformed E. coli could express and secret amylase. Compared with the original B.subtilis, the amylase activitives from transformed E.coli were 1.0376 U/mL in LB medium and 1.3610 U/mL in a special mineral medium, which were increased by 46 and 286 folds, respectively.
出处
《畜牧与兽医》
北大核心
2005年第6期7-9,共3页
Animal Husbandry & Veterinary Medicine
关键词
淀粉酶基因
转基因
大肠杆菌
枯草杆菌
amylase gene
transformation
Escherichia coli
Bacillus subtilis
