摘要
以牛乳素为目的基因,构建含有组氨酸标签、人生长激素信号肽的真核表达载体,为利用牛乳素在活体动物及培养细胞中的表达提供基因材料。用人工合成hGH-Lacf B的序列与T载体连接,用Xho I和BamH I酶切后与相应酶切处理的pVAX1连接,取阳性克隆质粒,用EcoR I和Xho I酶切回收含hGH的pVAX1载体,再与经过PCR扩增的his-Lacf B连接,酶切鉴定及测序。DNA序列分析表明,携带hGH、6×his和目的基因Lacf B已成功连接到真核表达载体pVAX1中。
To provide the eukaryotic expression vector carrying 6x histidine-tag and human growth hormone in expremental animal and cell culture, according to the published nucleotide sequence of the lactofericin B and human growth hormone, synthesis sequence was ligated with pMT-18 vector. The product was digested with Xho I and BamH I,and then cloned into the plasmid pVAX1.The recombinant plasmid was digested with EcoR I and Xho I, got the pVAX-hGH,and ligating with the gene encoding for his-lacfB and amplified using PCR technique. The eukaryotic expression vector carrying hGH.6x histidine-tag and lactofericin B had been established and verified by partial DNA sequence analysis.
出处
《湖北农业科学》
北大核心
2008年第5期501-502,570,共3页
Hubei Agricultural Sciences
基金
山东信得科技股份有限公司自主研发课题
关键词
牛乳素
组氨酸标签
人生长激素信号肽
真核表达
lactoferricin B
histidine tag
human growth hormone
eukaryotic expression