摘要： 克隆天祝白牦牛脱嘌呤嘧啶核酸内切酶基因(APEX-1)全长cDNA序列,为研究天祝牦牛高原抗逆性机制提供理论基础。根据已知哺乳动物APEX-1基因cDNA序列同源性,设计引物以及5’和3’特异性引物,运用cDNA末端快速扩增(RACE)技术获得天祝白牦牛APEX-1基因全长cDNA序列。扩增获得天祝白牦牛APEX-1基因全长cDNA序列为1 391 bp(GenBank登录号:KF611690),ORF长957 bp,编码318个氨基酸。氨基酸序列分析显示天祝白牦牛APEX-1基因编码氨基酸序列与大多数哺乳动物相似,但有7个氨基酸突变。同源建模预测,显示天祝白牦牛APEX-1蛋白由8个α-螺旋和11个β-折叠片形成一个呈三明治的α/β球蛋白。天祝白牦牛APEX-1基因的成功克隆为进一步研究该基因的功能奠定了基础。
关键词:
天祝白牦牛,
脱嘌呤嘧啶核酸内切酶,
cDNA末端快速扩增,
序列分析
Abstract: To clone Tianzhu White Yak apurinic /apyrimidinic endonuclease-1 (APEX-1 ) full length cDNA, thus provide a theoretical basis to study its mechanisms in stress resistance. Primers of the APEX-1 gene and its RACE primers were designed according to the alignment of known cDNA sequences of mammals, to obtain the full length cDNA of APEX-1 by PCR and RACE technique. The result showed that the full length of APEX-1 cDNA was 1 391 bp(GenBank accession No. KF611690 ), with an ORF 957 bp in length, which encoded 318 amino acids. The amino sequence analysis indicated that APEX-1 gene is highly homologous in most mammals. However, there are seven differences in amino sequence compared with other mammals. Homogenous 3D model of APEX-1 obtained by predict shows that APEX-1 protein is globular α /β rich protein, containing 11 β-sheets at the core which is surroun- ded by 8 helical structure. The full length cDNA of 1 391 bp APEX-1 gene was successfully cloned, which laid a ba- sis for further studying of its roles in White Yak stress resistance.
Key words:
Tianzhu White Yak,
APEX-1 gene,
Rapid amplification of cDNA ends (RACE),
Sequence analysis
中图分类号:
李亚兰, 张全伟, 王雪莹, 马友记, 张勇, 赵兴绪. 天祝白牦牛APEX-1基因的克隆及生物信息学分析[J]. 华北农学报, 2014, 29(2): 51-55. doi: 10.7668/hbnxb.2014.02.009.
LI Ya-lan, ZHANG Quan-wei, WANG Xue-ying, MA You-ji, ZHANG Yong, ZHAO Xing-xu. Cloning and Bioinformatics Analysis of APEX-1 Gene in Tianzhu White Yak[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(2): 51-55. doi: 10.7668/hbnxb.2014.02.009.