CAJ | 학술논문

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cD-NA序列为1391 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A-PEX-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。
극륭천축백모우탈표령밀정핵산내절매기인( APEX-1)전장cDNA서렬,위연구천축모우고원항역성궤제제공이론기출。근거이지포유동물APEX-1기인cDNA서렬동원성,설계인물이급5′화3′특이성인물,운용cDNA말단쾌속확증( RACE)기술획득천축백모우APEX-1기인전장cDNA서렬。확증획득천축백모우APEX-1기인전장cD-NA서렬위1391 bp(GenBank등록호：KF611690),ORF장957 bp,편마318개안기산。안기산서렬분석현시천축백모우APEX-1기인편마안기산서렬여대다수포유동물상사,단유7개안기산돌변。동원건모예측,현시천축백모우A-PEX-1단백유8개α-라선화11개β-절첩편형성일개정삼명치적α/β구단백。천축백모우APEX-1기인적성공극륭위진일보연구해기인적공능전정료기출。
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 globularα/βrich protein,containing 11β-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.