马尾松毛虫腺苷酸转移酶基因的克隆与分析 - 生物通.doc

马尾松毛虫腺苷酸转移酶基因的克隆与分析 Cloning and Primary Study of Adenine Nucleotide Translocase Gene in Dendrolimus puntatus WENG Hong-biao1, NIU Bao-long1, HE Li-hua1, SHEN Wei-feng1, WANG Shou-xian1,2, WU Zhong-liang3, JIANG Ping3, MENG Zhi-qi1 (1Laboratory of Entomo-Molecular Biology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021; 2Zhejiang Forestry University, Lin’an 311300, Zhejiang; 3Forest Pest Control and Quarantine Burean of Zhejiang Province, Hangzhou 310020) Abstract: 【Objective】 A complementary DNA clone encoding the adenine nucleotide translocases in Dendrolimus puntatus has been identified. 【Method】 RACE (Rapid Amplification of cDNA Ends) method was used to get the complete cDNA sequence. The sequence was analyzed with software. 【Result】The full length of adenine nucleotide translocases cDNA was cloned. (GenBank accession number: EF194157). he sequence contains a single open reading frame that encodes a polypeptide 300 amino acids in length. This polypeptide shows extensive similarities to the known ekaryotic translocase polypeptides, the similarity being greatest (up to 80% identity) to the Lepidoptera ADP/ATP translocases. 【Conclusion】Adenine nucleotide translocases of Dendrolimus punctatus has homology with that of other insects. Southern blot analysis suggests that a single copy of this gene is present per haploid genome. Key words: Dendrolimus punctatus; Adenine nucleotide translocases; Clone 0 引言 【研究意义】线粒体是细胞内主要的ATP生产中心，在细胞凋亡、信号转导的发生过程中起着非常重要的作用。腺苷酸转移酶denine nucleotide translocase，ANT是真核细胞的线粒体内膜上的转运蛋白家族成员[1]，在线粒体内膜上分布最为丰富，约占细胞线粒体内膜总蛋白的10%[1]，作为重要代谢物的载体，参与线粒体的各种活动[3]。【前人研究进展】ANT蛋白由两个完全相同，分别含一个ATP/ADP结合位点的亚基组成，通过其构象的变化，实现细胞质中ADP和线粒体内ATP间的跨膜交换[2,4,5]，是生物体内能量产生与能量消耗的重要连接[6]，生物体大约1/2～1/3的基础电子传递由ANT催化，是保持细胞内能量代谢的关键成分[7,8]。ANT虽非组成线粒体内膜上通透转换孔洞的必要成分，但与内质网与线粒体之间的Ca2+离子转移有关，参与线粒体Ca2+离子内环境平衡[9,10]。近来的研究表明，ANT在细胞凋亡诱导过程中有重要作用[11,12]，是各种细胞凋亡因子或凋亡抑制因子的作用靶位[1316]，细胞内具氧化活性的细胞凋亡诱导因子通过氧化ANT蛋白的巯基基团，诱导线粒体肿胀和线粒体内的SOD、细胞色素C的释放[17]，而细胞色素C则可激活细胞凋亡代谢阶梯[4,18]。大多数真核生物至少有2个不同的