Molecular Cloning, Expression, and Chromosomal Mapping of the Porcine CDC-2-Like Kinase 1 (CLK1) Gene

Molecular Cloning, Expression, and Chromosomal Mapping of the Porcine CDC-2-Like Kinase 1 (CLK1) Gene
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猪 CDC-2 样激酶 1 (CLK1) 基因的分子克隆、表达和染色体定位

DOI:
10.1007/s10528-009-9226-6
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发表时间:
2009-02
期刊:
Biochem Genet
影响因子:
--
通讯作者:
李加琪
李加琪
中科院分区:
其他
文献类型:
--
作者:
李加琪

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CDC-2样激酶1(CLK 1)在调节前RNA剪接和转录后基因表达中起关键作用。通过电子克隆、RT-PCR和RACE技术,鉴定了猪CLK 1基因的两种不同转录本,即全长CLK 1和截短型CLK 1(CLK 1 T)。该基因全长cDNA序列为1771 bp,开放阅读框为1455 bp,编码484个氨基酸。该基因全长1680 bp,开放阅读框414 bp,编码137个氨基酸。利用中国二花脸猪的细菌人工染色体克隆,分析了猪CLK 1基因组结构和序列,该基因包含13个外显子和12个内含子,全长约9 kb。RT-PCR结果显示,全长和截短剪接形式在猪心脏,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、小肠和肾脏的组织中,全长剪接形式的表达水平远高于截短形式。使用IMpRH面板将CLK 1基因物理地分配到微卫星标记SW 1316和SW 2083之间的SSC 15。
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 porcineCLK1gene, known as full-lengthCLK1and truncatedCLK1(CLK1T), were identified byin silicocloning, RT-PCR, and RACE. The entire cDNA sequence of full-lengthCLK1was 1771 bp, containing a 1455 bp ORF encoding a deduced protein of 484 amino acids. The complete cDNA sequence ofCLK1Tis 1680 bp, containing a 414 bp ORF encoding a deduced protein of 137 amino acids. The genomic structure and sequence of porcineCLK1were 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. TheCLK1gene was physically assigned to SSC 15 between microsatellite markersSW1316andSW2083using the IMpRH panels.
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