中华卵索线虫β-actin基因的克隆与分析

Cloning and Analysis ofβ-actin Gene in Ovomermis sinensis

利用RACE技术克隆中华卵索线虫Ovomermis sinensisβ-actin基因,RT-PCR方法检测该基因在中华卵索线虫不同发育时期的表达情况。结果首次获得了中华卵索线虫β-actin基因全长cDNA序列。该基因cDNA全长1636bp,其中ORF长1131bp,编码376个氨基酸;5′UTR长137bp;3′UTR长367bp。由该cDNA推导的蛋白质序列与秀丽隐杆线虫Caenorhabditis elegans、人类等物种的β-actin蛋白序列相似性均在95%以上。系统进化树显示其系统发育关系与传统的分类关系基本一致。β-actin基因在不同发育时期的线虫中持续恒量表达。由此推断中华卵索线虫β-actin基因具有高度保守性,可以作为生物物种进化的分子标志,且能较好反映物种间的系统发生关系,是量化实验的理想内标参照。

RT-PCR and RACE（rapid amplification of cDNA Ends） methods were used to cloneβ-actin gene in Ovomermis sinensis.The nucleotide sequence and the amino acid sequences were analyzed by related programs.The expression pattern ofβ-actin gene was also analyzed by RT-PCR.The full length ofβ-actin gene cDNA in O.sinensis was cloned for the first time.Theβ-actin gene is 1,636 bp in length, including an ORF of 1,131 bp encoding a protein of 255 amino acids,its 5＇-UTR and 3＇-UTR is 137 bp, 367 bp long,respectively.The identities of O.sinensisβ-actin are above 95%,compared with otherβ-actins. The phylogenetic analysis was consistent with traditional classification,suggesting that the reported O.sinensisβ-actin belongs to theβ-actin family.The expression of the clonedβ-actin gene was also consistent well in the different developmental stages of O.sinensis.Theβ-actin gene of O.sinensis is highly conserved,and can be used for the molecular markers of the biological evolution.It is also a good reference gene for semi-quantitative analysis.