In vitro expression of human cytomegalovirus UL148 RNA and prediction of its functional sites
10.3760/cma.j.issn.0254-5101.2018.02.003
- VernacularTitle:人巨细胞病毒临床病毒株UL148基因的体外表达及功能位点预测
- Author:
Jingjing HU
1
;
Yuanbin WU
;
Qiqi TAN
;
Haihao SU
;
Juncai DING
;
Yuanyuan GUO
;
Binhua XIE
;
Lijun CAI
;
Mengjie GUO
;
Bo WANG
Author Information
1. 广东省妇幼保健院儿科
- Keywords:
Human cytomegalovirus;
Clinical virus strain;
UL148;
Signal transduction
- From:
Chinese Journal of Microbiology and Immunology
2018;38(2):94-97
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.
