CAJ | 학술논문

目的构建CD151两种突变体,为研究CD151的功能及CD151-整合素复合体的功能奠定基础.方法以pAAV-CD151质粒为模板,采用定点突变技术,构建整合素α3β1/α6β1结合缺陷QRD突变体(pAAV-CD151-QRD-AAA~(194-196)突变体)和CD151囊泡运输基序缺陷的ARSA突变体(pAAV-CD151-YRSL-ARSA~(245-248)突变体),并包装两种突变体的重组腺相关病毒.结果经过测序鉴定成功构建了CD151的QRD和ARSA突变体,亦成功包装了重组腺相关病毒,病毒滴度符合本实验要求.结论成功构建CD151两种突变体并包装成重组腺相关病毒,为进一步研究CD151的功能和作用机制奠定了基础.
목적 구건CD151량충돌변체,위연구CD151적공능급CD151-정합소복합체적공능전정기출.방법 이pAAV-CD151질립위모판,채용정점돌변기술,구건정합소α3β1/α6β1결합결함QRD돌변체(pAAV-CD151-QRD-AAA~(194-196)돌변체)화CD151낭포운수기서결함적ARSA돌변체(pAAV-CD151-YRSL-ARSA~(245-248)돌변체),병포장량충돌변체적중조선상관병독.결과 경과측서감정성공구건료CD151적QRD화ARSA돌변체,역성공포장료중조선상관병독,병독적도부합본실험요구.결론 성공구건CD151량충돌변체병포장성중조선상관병독,위진일보연구CD151적공능화작용궤제전정료기출.
Objective To establish a basic method to construct two kinds of CD151 mutants for studying the function of CD151 and CD151-integrin ICAP1 compound body.Methods Taking pAAV-CD151 plasmid as the template,two CD151 mutants.pAAV-CD151-QRDAAA~(194-196) and pAAV-CD151-YRSL-ARSA~(245-248),were constructed by applying the technology of oriented direction mutation.Results Two kinds of CD151 mutants were successfully constructed,and the results were confirmed by sequencing.The recombinant adeno-associated viruses of CD151 mutants were successfully packaged,and the virus titers satisfied the experiment request.Conclusion Two kinds of CD151 mutants,pAAV-CD151-QRD-AAA~(194-198) and pAAV-CD151-YRSL-ARSA~(245-248),were successfully constructed,and the recombinant adeno-associated viruses of CD151 mutants were successfully packaged,which laid the foundation for further study on the CD151 functions and molecular mechanism.