Unconserved C terminal of pUL76 in human ctomegalovirus elicits aggresome formation
- VernacularTitle:人巨细胞病毒pUL76蛋白非保守C端与蛋白聚集体形成的研究
- Author:
Wenchang ZHANG
;
Jingxian CHEN
;
Mingli WANG
- Publication Type:Journal Article
- Keywords:
human cytomegalovirus;
UL76;
protein aggresome
- From:
Acta Universitatis Medicinalis Anhui
2015;(4):411-414,415
- CountryChina
- Language:Chinese
-
Abstract:
Objective To define the nuclear agrresome formation is determinated by which part of the UL76 of
HCMV. Full-length, conserved N terminal and unconserved C terminal of pUL76 were constructed to eukaryotic ex-pression plasmid pEGFP-N1 . Methods Primers were designed to amplify full-length and different part of pUL76 according to standard sequence of HCMV AD169 which had been submitted to GenBank(FJ527563. 1). These frag-ments were constructed to eukaryotic expression plasmid pEGFP-N1 . The recombinant plasmids were designated pEGFP-UL76,pEGFP-UL76N, pEGFP-UL76C respectively. Double digestion and sequencing were performed to verify the accuracy of recombinant plasmids construction. Empty vector and three recombinant plasmids were transi-ent transfected to HELF and HepG-2 cells respectively. Reverse transcriptation PCR and Western blot were per-formed to detect the RNA and protein expression level respectively. Different nuclear aggresome formations were visualized with an Olympus fluorescence microscopy. Results pEGFP-N1 and pEGFP-UL76N were unable to in-duce nuclear aggresome formation, whereas pEGFP-UL76 and pEGFP-UL76C were able to elicit nuclear aggresome formation. Conclusion The unconserved C terminal of pUL76 is sufficient to induce nuclear aggresome formation.
