猪Cdk2基因的克隆及其功能研究

doi:10.11843/j.issn.0366-6964.2013.10.003

Abstract

Abstract:

This study was aimed to clone the porcine cyclin-dependent kinase 2 (Cdk2) gene and investigate the function of CDK2 encoded by Cdk2. In this study, porcine Cdk2 cDNA was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Bioinformatic approaches were used to analyze the nucleotide and amimo acid sequences as well as the bio-function of CDK2. Semi-quantitative RT-PCR was used to analyze the tissue expression profile of Cdk2 mRNA, confocal microscopy was used to observe the subcellular localization of CDK2. Overexpression and shRNA techniques were both employed to investigate the role of CDK2 in cell cycle and cell proliferation regulation. Results showed that Cdk2 gene (GenBank accession no. JX967576) contained an 897 bp open reading frame and shared 94.2%, 94.0%, 93.8%, 93.4%, 91.8%, 91.0%, 90.6% and 89.9% identity with those of sheep, cattle, goat, human, golden hamster, house mouse, Chinese hamster and norway rat, respectively. Porcine Cdk2 gene coded 298 aa, with a molecular weight of 34 ku. The mRNA transcripts of porcine Cdk2 were found in 10 different porcine tissues by semi-quantative RT-PCR. Porcine CDK2 was localized in both the nucleus and cytoplasm and degradated through proteasomal pathway. Porcine CDK2 overexpression induced a significant decrease in proportion of cell at S-phase and G2/M-phase arrest, but no changes at G0/G1-phase. In contrast, when the CDK2 expression was down-regulated by shRNA, a significant decrease in proportion of cell at S-phase and G0/G1-phase arrest was observed, without changes at G2/M-phase. The porcine Cdk2 gene was successfully cloned and its function was also investigated.

CLC Number:

S828
S813.3

TANG Qing-hai, ZHANG Hui, WEI Yan-wu, LIU Chang-ming. Cloning and Functional Analysis of Porcine Cdk2 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1522-1531.

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Cloning and Functional Analysis of Porcine Cdk2 Gene

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