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猪细胞周期蛋白依赖性激酶2样激酶1(CLK1)基因的分子克隆、表达及染色体定位

Molecular cloning, expression, and chromosomal mapping of the porcine CDC-2-like kinase 1 (CLK1) gene.

作者信息

Xie Shui Hua, Li Jia Qi, Chen Yao Sheng, Gao Ping, Zhang Hao, Li Ze Yue

机构信息

College of Animal Science, South China Agricultural University, Guangzhou, People's Republic of China.

出版信息

Biochem Genet. 2009 Apr;47(3-4):266-75. doi: 10.1007/s10528-009-9226-6. Epub 2009 Feb 5.

Abstract

CDC-2-like kinase 1 (CLK1) plays a critical role in regulating pre-RNA splicing and post-transcriptional gene expression. Two distinct transcripts of the porcine CLK1 gene, known as full-length CLK1 and truncated CLK1 (CLK1 ( T )), were identified by in silico cloning, RT-PCR, and RACE. The entire cDNA sequence of full-length CLK1 was 1771 bp, containing a 1455 bp ORF encoding a deduced protein of 484 amino acids. The complete cDNA sequence of CLK1 ( T ) is 1680 bp, containing a 414 bp ORF encoding a deduced protein of 137 amino acids. The genomic structure and sequence of porcine CLK1 were analyzed using a bacterial artificial chromosome clone of a Chinese Erhualian pig, with 13 exons and 12 introns spanning approximately 9 kb. RT-PCR revealed that the full-length and truncated splice forms were expressed at equivalent levels in pig heart, fat, liver, spleen, and lymph tissues. The full-length splice form was expressed at a much higher level than the truncated form in tissues of the pig cerebrum, longissimus dorsi, small intestine, and kidney. The CLK1 gene was physically assigned to SSC 15 between microsatellite markers SW1316 and SW2083 using the IMpRH panels.

摘要

CDC-2样激酶1(CLK1)在调节前体RNA剪接和转录后基因表达中起关键作用。通过电子克隆、逆转录-聚合酶链反应(RT-PCR)和cDNA末端快速扩增(RACE),鉴定出猪CLK1基因的两种不同转录本,即全长CLK1和截短型CLK1(CLK1(T))。全长CLK1的完整cDNA序列为1771 bp,包含一个1455 bp的开放阅读框(ORF),编码一个推导的484个氨基酸的蛋白质。CLK1(T)的完整cDNA序列为1680 bp,包含一个414 bp的ORF,编码一个推导的137个氨基酸的蛋白质。利用中国二花脸猪的细菌人工染色体克隆分析了猪CLK1的基因组结构和序列,其有13个外显子和12个内含子,跨度约9 kb。RT-PCR显示,全长和截短型剪接形式在猪心脏、脂肪、肝脏、脾脏和淋巴组织中的表达水平相当。在猪大脑、背最长肌、小肠和肾脏组织中,全长剪接形式的表达水平远高于截短型。利用IMpRH板将CLK1基因物理定位到15号猪染色体上微卫星标记SW1316和SW2083之间。

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