猪TRIM11基因克隆及其促进猪伪狂犬病毒在体外增殖的研究

doi:10.11843/j.issn.0366-6964.2016.06.020

Abstract

Abstract:

In order to preliminary study the immunologic functions of porcine TRIM11，the cDNA of the target gene was amplified from the cDNA of porcine submandibular lymph node，the protein structure of porcine TRIM11 was predicted and its tissue expression profile was analysed，the cloned porcine TRIM11 cDNA was connected into PiggyBac (PB) transposon vector，then the recombinant vector (PB-TRIM11) was transfected to PK15 cells to establish stably transfected PK15 cell line.The results showed that the coding length of the cloned porcine TRIM11 was 1 407 bp and the sequence code 468 amino acids.Porcine TRIM11 protein was characterized by the presence of the tripartite motif.Tissue transcription profile analysis showed that，the transcription of TRIM11 was higher in fat，lung，whereas the transcription of porcine TRIM11 was lower in heart，ileum，recutum，duodenum.DNA sequence analysis of TRIM11 showed that the cloned TRIM11 cDNA was consistent with sequence from GenBank and the recombinant vector (PB-TRIM11) was constructed.The transfected PK15 cell line stably expressing TRIM11 was established successfully and the transcription of TRIM11 was identified by qRT-PCR.PK15 cell line that stably express porcine TRIM11 and control cells were infected with pseudorabies virus (PRV)，respectively，then the cell supernatants were harvested at various time points after infection and the virus titers were detected.The results showed that the viral TCID₅₀ of the TRIM11 over-expression group was higher than the control all the time.It indicates that the TRIM11 over-expression cells might be more susceptible to PRV.The study provides the experimental basis for the role of porcine TRIM11 in the innate immunity system in the future.

CLC Number:

SONG Shuang，ZENG Lei，LU Shao-fang，GUO Zhen-zhen，LU Wei-fei，HAN Li-qiang，WANG Jiang，WANG Yue-ying，CHU Bei-bei，YANG Guo-yu. Molecular Cloning of Porcine TRIM11 Gene and Its Promotion Effect on Pseudorabies Virus Replication in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(6): 1239-1246.

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Molecular Cloning of Porcine TRIM11 Gene and Its Promotion Effect on Pseudorabies Virus Replication in vitro

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