秦川牛CAP2基因CDS区克隆及表达谱分析

doi:10.12160/j.issn.1672-5190.2022.03.003

Abstract

Abstract: [Objective] To clone the coding region sequence （CDS） of CAP2 gene in Qinchuan bovine, and to characterize the expression profiling of this gene in various tissues and during the primary adipocyte differentiation. [Method] The CDS region of CAP2 gene in Qinchuan bovine was amplified by RT-PCR assay, and the CAP2 protein was analyzed using online bioinformatical tools such as ProtPram, TMpred, and ProtFun 2.1 Server. The gene expression levels of PPARγ and FABP4 were detected using qPCR assay and oil red O staining method to construct a bovine primary adipocyte induced differentiation system. Moreover, the expression levels of CAP2 gene in seven tissues （heart, liver, spleen, lung, kidney, muscle, and back fat） as well as during the differentiation process of primary adipocyte （0-10 days） in Qinchuan bovine were assessed using qPCR assay. [Result] The CDS region of CAP2 gene in Qinchuan bovine was 1 461 bp and encoded 486 amino acids. Its amino acid sequence mainly composed of random coil and α-helix. Extremely significantly （P<0.01） higher expression level of CAP2 gene was observed in back fat in comparison to the other tested tissues. During the differentiation process of primary adipocyte, the expression levels of PPARγ and FABP4 genes increased gradually and peaked on day 10, which were extremely significantly （P<0.01） higher compared to day 0. The induced differentiation group of primary adipocyte had a much larger lipid droplet accumulation than the control group. Furthermore, the expression level of CAP2 gene increased gradually with the prolongation of induced differentiation duration, and peaked on day 10 as well, which were extremely significantly （P<0.001） higher compared to day 0. [Conclusion] The 1 461 bp full-length CDS of CAP2 gene in Qinchuan bovine has been successfully cloned. The CAP2 gene is highly expressed in adipose tissue and may involves in adipogenesis and differentiation. As a result, the CAP2 gene is assumed to be a transcription factor that promotes adipogenic differentiation, plays a key role in regulating the status of bovine adipocyte, and could be a candidate gene associated with meat quality trait of Qinchuan bovine. These findings provide preliminary information for further revealing the function of bovine CAP2 gene.

Key words: Qinchuan bovine, CAP2 gene, adipocyte, expression pattern, bioinformatic analysis

CLC Number:

S823.2

GAO Yu-hong, HU Chun-li, MA Yan-fen, WANG Shu-zhe, MA Yun. Cloning and Expression Profiling of CDS Region of CAP2 Gene in Qinchuan Bovine[J]. Animal Husbandry and Feed Science, 2022, 43(3): 16-24.

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URL: https://journal30.magtechjournal.com/xmysl/EN/10.12160/j.issn.1672-5190.2022.03.003

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Cloning and Expression Profiling of CDS Region of CAP2 Gene in Qinchuan Bovine

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