兴凯湖翘嘴鲌(Culter alburnus)LPL基因克隆及其表达

Cloning and expression of LPL gene of Culter alburnus in Xingkai Lake

采用RT-PCR和RACE技术,进行兴凯湖翘嘴鲌LPL基因的克隆与表达分析,旨在为其脂肪性状的研究奠定基础.根据Genbank中已知的鲤科鱼类LPL基因序列设计引物,克隆出兴凯湖翘嘴鲌(Culter alburnus)脂蛋白酯酶(idella lipoprotein lipase,LPL)的c DNA序列;将其提交到Genbank,获得基因登陆号为KC166 231;兴凯湖翘嘴鲌LPL基因c DNA序列共1 947 bp,基因编码区序列共1 524 bp,编码一个终止密码子和507个氨基酸;通过Blast与Genbank上发表的其他物种的CDS区进行比对发现,与草鱼(Ctenopharyngodon)的同源性达到98%.采用荧光定量PCR(Real-time PCR)技术检测了兴凯湖翘嘴鲌野生群体和养殖群体在不同年龄、不同组织中LPL的表达量.结果表明,随年龄的增长野生群体与养殖群体LPL基因相对表达量差异增大,4龄养殖群体LPL基因相对表达量是同期野生群体的17.54倍,二者差异显著(P<0.05);LPL基因在心脏当中表达水平最高,脾和肝脏中的表达量次之.

In order to lay a foundation for the study of the characters of its fat, we carryed out the clo- ning of the LPL gene of Culter alburnus in Xingkai Lake and analyzed its expression using RT- PCR and RACE technology. According to the known LPL gene sequence designed primers of cyprinid fish in the Genbank, We cloned cDNA sequence of idelIa lipoprotein lipase of Culter alburnus and submitted it to the Genbank, getting Genbank accession no KC166 231. There were 1 947 bp cDNA sequences of the LPL gene of Culter alburnus in Xingkai Lake and 1 524 bp coding sequences. We coded one termi- nation codon and 507 amino acid. By comparing Blast with other species＇ CDS region published in the Genbank, we found its homology with Ctenopharyngodon reached 98%. We detected LPL expression quantity at different ages and in different tissues of cultured and wild populations of Culter alburnus in Xingkai Lake by using Real- time PCR and the results showed that, the differences of relative expres- sion of LPL gene were advanced with the growth of age of cultured and wild populations. The relative expression of LPL gene of cultured population at age four was 17.54 times of the homochronous wild population and they were diversity markedness （P 〈 0.05）. The expression level of LPL gene in heart was the highest, which of spleen and liver took the second place.