白绒山羊PPARγ基因RNA干扰慢病毒载体的构建及对肌内脂肪细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2016.04.005

Abstract

Abstract:

The cashmere-goat intramuscular adipocytes were used as experimental materials，and aimed to establish the stable intramuscular adipocyte line for the interference through target interrupting the peroxisome proliferation-activated receptor(PPARγ) gene，so as to explore the function of cashmere-goat PPARγ gene during the proliferation and differentiation processes of the intramuscular adipocytes.Lentivirural plasmids pack-aging system was adopted to establish a lentiviral vector for RNA interference of specific targeting White Cashmere-goat PPARγ gene，which was used to establish the intramuscular adipocyte line to stabilize and silence the PPARγ gene.Real-time quantitative and Western blot methods were used to detect the expression of PPARγ gene at different points in time in the interference group and control group，as well as the MTT and Oil Red O Staining methods were used to research the influence of cashmere-goat PPARγ gene on the proliferation and differentiation of intramuscular adipocytes.It was confirmed by sequencing that oligonucleotide chains containing PPARγ-shRNA lentivirus vector were inserted properly，its infection efficiency to intramuscular adipocytes was above 80%，lentivirus-mediated shRNA detected by fluorescence quantitative and Western blot could effectively reduce the expression of PPARγ gene，the reaction time interval between mRNA and protein level was 24 h.After silencing of PPARγ gene，the concentration of triglyceride within adipocytes was significantly lower than that in the control group；Contrarily，the cell proliferation ability in MTT test interference group was higher than that in control group.This study established cashmere-goat shRNA-PPARγ lentivirus interference vector，after successfully transfecting into the intramuscular adipocytes，it obviously inhibited the differentiation of adipocytes，and promoted the proliferation of intramuscular adipocytes，which lay foundation for further research the role of PPARγ gene in cashmere-goat metabolic pathways and in the fat deposition mechanism.

Key words: intramuscular adipocyte, RNA interference, PPARγ

CLC Number:

S827
S813.3

DU Chen，LI Jin-quan，CHEN Xiu-juan. Construction of Lentiviral RNAi Vector of PPARγ Gene in Cashmere Goat and Its Effect on Proliferation and Differentiation of Intramuscular Adipocytes[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 671-678.

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Construction of Lentiviral RNAi Vector of PPARγ Gene in Cashmere Goat and Its Effect on Proliferation and Differentiation of Intramuscular Adipocytes

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