为了研究猪瘟病毒E 2糖蛋白的立体结构及生物学特性,将增强型荧光蛋白基因(EGFP)和猪瘟兔化弱毒疫苗株(HCLV)E 2基因经PCR扩增后克隆至pB lueB acH is2A质粒,与杆状病毒DNA共转染后经PCR鉴定获得了含有EGFP和HCLV E 2融合基因(GFPTE 2)...为了研究猪瘟病毒E 2糖蛋白的立体结构及生物学特性,将增强型荧光蛋白基因(EGFP)和猪瘟兔化弱毒疫苗株(HCLV)E 2基因经PCR扩增后克隆至pB lueB acH is2A质粒,与杆状病毒DNA共转染后经PCR鉴定获得了含有EGFP和HCLV E 2融合基因(GFPTE 2)的重组杆状病毒rBACTE 2-339,并将其感染sf9细胞后在荧光显微镜下观察到了亮绿色荧光,说明融合基因已初步表达。展开更多
SPAP2,a transmembrane protein,is an Ig family receptor containing both ITIMs(immunoreceptor tyrosine-based inhibition motifs) and ITAMs(immunoreceptor tyrosine-based activation motifs).The extracellular portion of SPA...SPAP2,a transmembrane protein,is an Ig family receptor containing both ITIMs(immunoreceptor tyrosine-based inhibition motifs) and ITAMs(immunoreceptor tyrosine-based activation motifs).The extracellular portion of SPAP2 contains six immunoglobulin-like domains and its intracellular segment has two ITAMs and two ITIMs.Sequence alignment with the genomic database reveals that the SPAP2 gene contains 16 exons and is localized at chromosome 1q21.SPAP2 is consisted of 734 amino acids,and the intercellular portion of SPAP2 contains 137 amino acids. Tyrosine-phosphorylated SPAP2 is specifically associated with SH2 domain-containing tyrosine kinases and SH2 domain-containing tyrosine phosphatases,which lead to the initiation of signal transduction.SPAP2CT gene was amplified by PCR with SPAP2 full-length DNA as the template and cloned to the pBluescript Ⅱ KS vector.pGex-2T-SPAP2CT,the expression vector of dissoluble fusion protein,was constructed and transferred into E.coli of DE3-plysS.The fusion protein GST-SPAP2CT was expressed efficiently and purified by FFQ ion exchange chromatography and GSH affinity chromatography.The result indicates that we have constructed steady expression vector pGex-2T-SPAP2CT, which was expressed in E.coli.The molecular weight of the dissoluble fusion protein is 46 000,the productivity of GST-SPAP2CT protein is 10% and the purity is over 90% after the purification.展开更多
文摘为了研究猪瘟病毒E 2糖蛋白的立体结构及生物学特性,将增强型荧光蛋白基因(EGFP)和猪瘟兔化弱毒疫苗株(HCLV)E 2基因经PCR扩增后克隆至pB lueB acH is2A质粒,与杆状病毒DNA共转染后经PCR鉴定获得了含有EGFP和HCLV E 2融合基因(GFPTE 2)的重组杆状病毒rBACTE 2-339,并将其感染sf9细胞后在荧光显微镜下观察到了亮绿色荧光,说明融合基因已初步表达。
文摘SPAP2,a transmembrane protein,is an Ig family receptor containing both ITIMs(immunoreceptor tyrosine-based inhibition motifs) and ITAMs(immunoreceptor tyrosine-based activation motifs).The extracellular portion of SPAP2 contains six immunoglobulin-like domains and its intracellular segment has two ITAMs and two ITIMs.Sequence alignment with the genomic database reveals that the SPAP2 gene contains 16 exons and is localized at chromosome 1q21.SPAP2 is consisted of 734 amino acids,and the intercellular portion of SPAP2 contains 137 amino acids. Tyrosine-phosphorylated SPAP2 is specifically associated with SH2 domain-containing tyrosine kinases and SH2 domain-containing tyrosine phosphatases,which lead to the initiation of signal transduction.SPAP2CT gene was amplified by PCR with SPAP2 full-length DNA as the template and cloned to the pBluescript Ⅱ KS vector.pGex-2T-SPAP2CT,the expression vector of dissoluble fusion protein,was constructed and transferred into E.coli of DE3-plysS.The fusion protein GST-SPAP2CT was expressed efficiently and purified by FFQ ion exchange chromatography and GSH affinity chromatography.The result indicates that we have constructed steady expression vector pGex-2T-SPAP2CT, which was expressed in E.coli.The molecular weight of the dissoluble fusion protein is 46 000,the productivity of GST-SPAP2CT protein is 10% and the purity is over 90% after the purification.
