一种组织特异性兼可调控性慢病毒基因表达载体制造技术

本发明专利技术涉及一种慢病毒载体。具有组织特异性兼备可调控性双基因表达。其特征在于具有沿着５’至３’方向至少包括下列元件：ｐｏｌｙＡ６０尾巴、逆式四环素反应激活因子（ｒｔＴＡ２ｓ－Ｍ２）、血管内皮钙启动子（ＶＥｃａｄ），四环素调控性白蛋白融合启动子（ＴＲＥ／ａｌｂ）、荧光素酶基因（ｌｕｃｉｆｅｒａｓｅ）。本发明专利技术构建的载体的双基因表达盒采用反式表达的方式，最大限度减少了外源性核苷酸序列片段，使其可插入有容量限制的慢病毒载体，并对插入的荧光素酶基因不作限定，可以自由选择外源性目的基因与可调控性启动子搭配，该载体表达盒在不需要提高慢病毒量滴度转染细胞的同时既能保持外源性目的基因可控性表达且显著提高外源基因的表达效率。

Tissue specific and regulatory lentiviral vector for gene expression

The present invention relates to a lentiviral vector. It has both tissue-specific and regulatory dual gene expression. The utility model is characterized in that the 5 'to 3' direction at least comprises the following components: polyA60 tail, reverse tetracycline reaction activator (rtTA2s - M2), vascular endothelial cadherin promoter (VEcad), tetracycline regulated albumin fusion promoter (TRE / ALB) and luciferase gene (luciferase). Double gene vector constructed by the invention of the expression cassette by trans expression way, to minimize the exogenous nucleotide sequence fragment, so that it can be inserted into the lentiviral vector with limited capacity, and the luciferase gene into not limited, can choose the exogenous gene and inducible promoter collocation. The expression in the box does not need to raise the amount of virus titer slowly transfected cells also can maintain the exogenous gene controlled expression and significantly improve the expression efficiency of exogenous gene.

【技术实现步骤摘要】

【技术保护点】
１．一种慢病毒基因表达载体，其特征在于从５’－３’方向至少包括以下元件：ｉ．ＰｏｌｙＡ尾巴序列；ｉｉ．转录激活因子编码序列；ｉｉｉ．对应于组织特异性启动子序列；ｉｖ．对应于药物和转录激活因子的可调控性启动子；ｖ．包括编码至少一种目的外源基因序列。

【技术特征摘要】

【专利技术属性】
技术研发人员：杨光华，张文炜，王华，
申请(专利权)人：上海比昂生物医药科技有限公司，
类型：发明
国别省市：31[]