Abstract：[Objective]To construct pLVX‐IRES‐TDtomat‐GSK‐3βlentiviral expression vectors and infect murine bone marrow DC2 .4 cell line for establishing DC2 .4 cell line with an over‐expression of murine GSK‐3β.[Methods]Coding sequence in murine GSK‐3β cDNA was amplified by reverse transcription‐polymerase chain reaction (RT‐PCR) assay and recombined into pLVX‐IRES‐TDtomat plasmid .After confirming with re‐striction endonucleases and sequencing ,the recombinant plasmid was transfected into 293T cells with Lipo‐fectamine 2000 and then packaged into lentivirus particles .DC2 .4 cell line was infected by lentivirus particles . There were 3 groups of DC2 .4 cell line control ,non‐virus control and GSK‐3βlentivirus .The expression lev‐els of GSK‐3βamong three groups were detected by real‐time PCR and Western blot .[Results]Restriction en‐donuclease assay and sequence analysis verified the successful construction of recombinant vector pLVX‐IRES‐TDtomat‐GSK‐3β.And it was expressed highly in packaging cell 293T and the titer of recombinant lentivirus particles was 1 .26 × 108 TU/mL .GSK‐3βexpression level increased more in GSK‐3βlentivirus group than that in DC2 .4 cell line control and non‐virus control at the levels of mRNA and protein .[Conclusion]GSK‐3βlenti‐viral recombinant vector is successfully constructed and expressed stably in DC2 .4 cell line .The study may provide rationales for further elucidating the immunological functions of DC2 .4 .
基金资助:国家自然科学基金(81102293)
引用本文:
包杰%熊石龙. 小鼠GSK-3β慢病毒表达载体的构建及其在骨髓树突状细胞DC2．4中的表达[J]. 医学临床研究, 2015, 32(1): 13-15,16.
BAO Jie%XIONG Shi-long. Construction of Murine GSK3βLentiviral Expression Vector and its Expression in DC2 .4 Cell Li ne. JOURNAL OF CLINICAL RESEARCH, 2015, 32(1): 13-15,16.
2015, Vol. 32 
