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RP2是最近定位克隆的一个X连锁隐性的视网膜色素变性基因。用PCR法直接从人视网膜cDNA文库中扩增到包括人RP2基因编码区在内的cDNA ,先克隆至 pJLA5 0 3载体中 ,随后将RP2编码区基因亚克隆至表达载体 pPROEXHTa中 ,并进行了DNA序列分析。用IPTG诱导hRP2融合蛋白在大肠杆菌DH5α菌株中表达 ,3 0℃诱导 10h后重组蛋白质约占菌体总蛋白质的 7% ,而 3 7℃诱导 5h后重组蛋白质即占菌体总蛋白质的5 .6%。这些工作为纯化RP2蛋白及进一步研究RP2蛋白的理化性质、免疫组织定位和结构功能打下了一定的基础
RP2 is an X-linked recessive retinitis pigmentosa gene of the most recently located clone. PCR directly from human retinal cDNA library was amplified to include human RP2 gene coding region cDNA, first cloned into pJLA5 0 3 vector, then the RP2 coding region gene was subcloned into the expression vector pPROEXHTa, and carried out DNA sequence analysis. The hRP2 fusion protein was induced by IPTG in E. coli DH5α. The recombinant protein accounted for about 7% of the total bacterial protein after 10 h of induction at 30 ℃, while the recombinant protein accounted for 5% of the total bacterial protein 5 h after induction at 37 ℃. 6%. These work lay a solid foundation for purifying RP2 protein and further studying the physicochemical properties, immunohistochemical localization and structural function of RP2 protein