外来入侵生物福寿螺β-actin基因片段的克隆及表达分析

Cloning and expression analysis of cDNA fragments of β-actin protein from the golden apple snail,Pomacea canaliculata

采用RT-PCR的方法首次对入侵我国福寿螺的β-actin基因片段进行扩增、克隆和序列测定,并通过氨基酸序列分析推导与其他相关物种的亲缘关系.结果表明,获得的福寿螺β-actin基因cDNA片段大小为840bp,核酸编码有280个氨基酸序列,与其他物种具有高度的同源性.通过邻接法(NJ)构建的系统发育树显示,福寿螺首先与软体动物聚为一簇,然后与节肢动物聚为一簇,再与脊椎动物聚为一簇.此外,RT-PCR结果显示,β-actin基因在福寿螺的鳃、消化腺、肾和足部肌肉等4个组织内的表达基本一致,具有良好的稳定性.

Two cDNA fragment of β-actin gene in Pomacea canaliculata were amplified with a pair of degenerated digonudeotide primers by RT-PCR,and the length was 840 bp encoding 280 amino acids.Blast searching in NCBI showed that the deduced amino acids sequence was highly similar to that of other known related species.A phylogenetic tree analysis based on NJ(Neighbor-Joining) is shown that P.canaliculata belongs to mollusca.In addition,relative expression of β-actin mRNAs in the gill,digestive gland,kidney and foot muscle of snail are keeping constant by semi-quantitative RT-PCR.