Question

摘要翻译

目的：构建人α-防御素HNP-2大肠杆菌表达载体并转化大肠杆菌DH5α。方法：利用PCR技术从质粒pcDNA3.1(+)扩增人α-防御素HNP-2基因，HindⅢ和EcoRⅠ分别酶切目的基因和载体pET-28（+），利用T4 DNA酶连接构建重组表达载体pET-28(+)/HNP-2，转化大肠杆菌DH5α，经快速PCR筛选和酶切鉴定检测转化成功后，琼脂糖回收重组表达载体pET-28(+)/HNP-2，转入大肠杆菌BL21（DE3），经IPTG诱导表达后对其表达产物进行SDS-PAGE。结果：酶切鉴定证实构建的载体为pET-28(+)/HNP-2，SDS-PAGE电泳显示HNP-2在大肠杆菌中得到了表达。结论：成功构建真核表达载体pET-28(+)/HNP-2，HNP-2在大肠杆菌中成功表达，为其进一步生物学研究奠定了基础。

Answer 1

Objective: To construct the human α-defensin HNP-2 E. coli expression vector and transformed into E. coli DH5α. Methods: PCR technology from the plasmid pcDNA3.1 (+) amplification of human α-defensin HNP-2 gene, Hind Ⅲ and EcoR Ⅰ digestion target gene and vector pET-28 (+), using T4 DNA ligase to construct recombinant expression vector pET-28 (+) / HNP-2, transformed into E. coli DH5α, transformed by the rapid PCR screening and restriction analysis detected after the success of the the agarose recovery recombinant expression vector pET-28 (+) / HNP-2, into the large intestine Bacillus BL21 (DE3), induced by IPTG its expression product SDS-PAGE. Results: Enzyme digestion confirmed the constructed vector pET-28 (+) / HNP-2, SDS-PAGE electrophoresis shows the HNP-2 expression in E. coli. Conclusion: We successfully constructed the eukaryotic expression vector pET-28 (+) / HNP-2, HNP-2 in E. coli successfully expressed, which laid the foundation for its further biological studies.