中文摘要：

以SmaI和SphI双酶切pMD18-T-CBHⅡ和以胸苷酸合成酶基因（thymidylate synthase，thyA）为选择压力的非抗生素抗性的穿梭表达载体pW425t，胶回收酸性纤维素酶CBHⅡ基因和载体大片段，并将纯化的CBHⅡ基因和表达载体pW425t大片段进行连接，构建出可以在乳酸菌与大肠杆菌之间穿梭表达的原核表达重组质粒pW425t-CBHⅡ。将pW425t-CBHⅡ转化至thyA基因缺陷型的乳酸杆菌感受态细胞中，通过质粒提取、酶切鉴定、PCR鉴定、测序分析和生长功能弥补筛选阳性克隆。SDS-PAGE分析，可见约49.6 ku的蛋白，并且刚果红染色显示重组乳酸杆菌可产生明显的水解圈。

英文摘要：

Recombinant plasmids pMD18-T-CBH Ⅱ and pW425t, prokaryotic expression shuttle vector between E. coli and Lactobacillus, were digested with Sinai and SphI enzymes respectively. The purified CBH Ⅱ gene was subcloned into the expression vector pW425t. Thus, the recombinant pW425t-CBH Ⅱ was constructed, and then was transformed into the competence thyA gene-mutant Lactobacillus DOMLaS107. Treated lysates of bacterium were loaded directly on SDS-PAGE, and approximately 49.6 kD protein was observed, and recombinant Lactobacillus can produced clear hydrolysis halos on the Congo-Red-CMC plate.