摘要
根据猪舒血管肠肽氨基酸序列推导、设计、合成了VIP基因。构建重组表达载体pPICZα A VIP。转化Pichiapastoris,在含 1 0 0 μg mlZeocin的YEPD平板上筛选。重组子经Ni NTA介质亲和层析纯化并计算VIP的表达量约为 1 2 5g L左右 ,纯化样品通过小分子质量电泳并与VIP标准样品迁移率对照进一步确定了舒血管肠肽已在巴斯德毕赤酵母中得到表达。
According to porcine Vasoactive Intestinal Peptide (VIP) amine acids sequence, the vip gene was designed and synthesized.Recombinant expression vector pPICZα A VIP was constructed and Pichia pastoris GS115was transformed by the recombinant plasmid.The anti Zeocin colonies were screened on YEPD plates containing 100μg/ml Zeocin.The vip gene has integrated into the genome DNA of Pichia pastoris GS115 by analysis of PCR and enzyme digest,the expressed VIP was purified by Ni NTA affinity chromatograph,the expressed amount of VIP was about 1.25g/L, the purified VIP has the same transfer rate with the standard VIP in the electrophoresis of Tricine SDS PAGE,this furtherly confirmed the expression of VIP in Pichia pastoris .
出处
《中国生物工程杂志》
CAS
CSCD
2004年第10期42-46,共5页
China Biotechnology
基金
广东省自然科学基金资助项目 (980 174)