山羊RPL26基因生物信息学分析及对肌内脂肪细胞分化的影响

doi:10.11843/j.issn.0366-6964.2022.04.007

Abstract

Abstract: The purpose of this study was to clone the sequence of goat RPL26 gene and explore its expression in tissues and its regulation on adipocyte differentiation of goat. The one-year old male Jianzhou da’er goat (with good healthy growth status, weight of 50 kg, n=3) was selected as the experimental object, the RPL26 sequence was cloned by RT-PCR, and the sequence of gene and protein were analyzed by bioinformatics; Goat tissue cDNA was used as template to construct tissue expression profiles by qPCR; The overexpression vector of RPL26 was constructed by double enzyme digestion method. RPL26 was overexpressed in goat intramuscular adipocytes by liposome transfection. The morphological changes were observed by Oil red O staining and Bodipy staining after induction. The overexpression efficiency and expression of adipogenesis related genes were detected by qPCR to clarify its effect on intramuscular adipocyte differentiation. The results showed that the cloned goat RPL26 sequence was 544 bp, the CDS region was 438 bp, and the coding protein was negatively charged unstable hydrophilic protein, which was mainly located in the nucleus. RPL26 was widely expressed in goat tissues, and highly expressed in abdominal subcutaneous fat and back subcutaneous fat, which were extremely significantly higher than that in other tissues (P<0.01). RPL26 was overexpressed in goat intramuscular adipocytes, and the lipid droplets in intramuscular adipocytes increased significantly, the OD value detected by Oil red O staining showed the same result; qPCR results showed that the expression level of Pref1 decreased significantly(P<0.01), the expression levels of adipocytes differentiation marker genes such as PPARγ, and lipid metabolism marker gene such as LPL showed an extremely significantly upregulated expression(P<0.01). The cloned goat RPL26 sequence is 544 bp, and the CDS region is 438 bp, its highest expression was in abdominal subcutaneous fat. RPL26 has a positive regulatory effect on the differentiation of goat intramuscular preadipocytes.

Key words: goat, ribosomal protein, clone, intramuscular adipocytes, cell differentiation

CLC Number:

S827.2

ZHANG Hao, WANG Yong, LI Yanyan, LUO Cheng, LI Xin, LI Zhixiong, ZHU Jiangjiang, LIN Yaqiu. Bioinformatics Analysis of Goat RPL26 Gene and Its Effect on Intramuscular Adipocyte Differentiation[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1064-1076.

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Bioinformatics Analysis of Goat RPL26 Gene and Its Effect on Intramuscular Adipocyte Differentiation

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