过表达MAT2A基因促进猪肌内脂肪细胞分化的研究

doi:10.16431/j.cnki.1671-7236.2018.09.010

Abstract

Abstract:

This study was aimed to explore the role of methionine adenosyltransferase 2A (MAT2A) in porcine intramuscular adipocyte differentiation by constructing the adenovirus mediated overexpression vector in vitro.The primers was designed according to the MAT2A gene mRNA sequence in GenBank (accession No.:NM_001167650.1),the total RNA sequence of porcine adipose tissue cell was extracted and reversed to get cDNA,which was as the template to proceed PCR amplification and connected to the pAdTrack-CMV adenovirus shuttle vector for sequencing identification,the recombinant plasmid was named pAd-MAT2A.pAd-MAT2A was digested by Pac Ⅰ and transfected into 293A cell.The expression level of MAT2A gene was detected by Real-time PCR,and the MAT2A protein was analyzed by Western blotting.The cells differentiated on the 8th day were used for Oil red O staining.The results showed that the shuttle vector (pAdTrack-CMV-MAT2A) was successfully constructed and homologous recombinated with backbone vector (pAdEasy-1).The virus titer was 1E+6 PFU/mL after transfecting the 293A cell with adenovirus vector pAd-MAT2A.Real-time PCR and Western blotting showed that the mRNA and protein levels of MAT2A gene were significantly upregulated by transfecting pAd-MAT2A in porcine intramuscular preadipocytes.Oil red O staining results showed that the overexpression of MAT2A gene promoted lipid accumulation in porcine intramuscular adipocytes.In conclusion,MAT2A gene was overexpressed in expression vector mediated by adenovirus and promoted the lipid increasement in porcine intramuscular adipocytes.

Key words: MAT2A gene; recombinant adenovirus; porcine; intramuscular preadipocyte

CLC Number:

ZHAO Cunzhen, WU Haigang, TANG Xuefeng, JIAO Fengchao, MA Yun, LIU Jicheng. Study on Overexpression of MAT2A Gene Promotes Porcine Intramuscular Preadipocyte Differentiation[J]. , 2018, 45(9): 2425-2432.

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Study on Overexpression of MAT2A Gene Promotes Porcine Intramuscular Preadipocyte Differentiation

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