RCAS(J)-Luciferase重组病毒拯救及其相关生物学功能鉴定

Rescue of recombinant RCAS(J)-Luciferase virus and its application in the study of subgroup J avian leukosis virus receptor

RCAS载体系列是基于A型罗斯肉瘤病毒(RSV-A)所改造而成的反转录病毒感染性克隆,转染易感细胞后,能够产生高滴度的RSV-A病毒。同时,RCAS载体可携带外源基因进入靶细胞,从而进行蛋白的高效表达。为了获取滴度高且易于检测的RSV,进行ALV-J受体方面的研究,本研究将RCAS(A)-GFP载体中RSV-A的囊膜蛋白基因替换为ALV-J的囊膜蛋白基因,并将绿色荧光蛋白报告基因(gfp)替换为海参荧光素酶(luciferase)报告基因,构建带有luciferase报告基因的囊膜蛋白为ALV-J囊膜蛋白的RSV重组病毒。经验证,本研究所构建的重组病毒,与传统的ALV-J株相比,具有复制能力强,病毒滴度高等优点,而且能够通过ALV-J特异性受体ch NHE1感染ALV-J非允许细胞系,有利于病毒感染初期的检测和定量,为病毒与其受体之间相互作用的研究奠定了基础。

RCAS vectors are retrovirus infectious clones based on Ross sarcoma virus A （RSV-A）, which are able to produce high titer virus. At the same time, the RCAS vectors carry exogenous genes into target cells, where the exogenous genes expressed efficiently. For obtaining RSV of high titers and easy detection, so as to contribute to the study of ALV-J receptor, the replaced the env gene and gfp reporter gene in RCAS （A）-GFP were replaced with the ALV-J env gene and the luciferase reporter gene, respectively. Then, a recombinant RSV expressing the envelop of ALV-J and luciferase was rescured which had higher replication ability to produce high virus titer, and the rescured virus was also able to infect ALV-J non permissive cells by interacting with ALV-J specific receptor, which helps the detection and quantification of virus in the early stage of infection and therefore lay the foundation for the related study of interaction between virus and its cellular receptor.