家蚕丝氨酸蛋白酶抑制剂基因serpin-6的克隆、序列分析和组织表达

Molecular cloning,sequence analysis and tissue expression of serpin-6 gene in Bombyx mori

本文利用RT-PCR和RACE方法,获得了家蚕Bombyxmori丝氨酸蛋白酶抑制剂基因(serpin)的开放读码框(ORF)和5′UTR序列。根据该基因与其他昆虫serpin的同源性关系将其命名为家蚕serpin-6基因,GenBank登录号EU159447。为了研究serpin-6与家蚕免疫反应的相关机理,对serpin-6基因的组织表达和免疫刺激反应进行了研究。结果表明该基因在家蚕的头部和生殖腺中mRNA表达量非常高,在中肠,脂肪体,丝腺和血淋巴表达量较低。用脂多糖(LPS)刺激5龄第3d家蚕后,serpin-6基因在脂肪体和血淋巴中都被显著诱导表达,且都在刺激6h后表达量最高。推测该基因在家蚕细胞免疫反应中起着一定的作用。

The ORF and the 5＇UTR sequences of serpin were cloned from Bombyx mori using RT-PCR and rapid amplification of cDNA ends （RACE） strategies. We named this gene as serpin-6 of B. mori, which was deposited in GenBank with the accession no. EU159447.. In order to study the mechanism of immunity of serpin-6 from B. mor/, we performed the tissue expression and detected the immunity response. The results of semi-quantitative RT-PCR showed that the mRNA of this gene was highly expressed in head and gonads, and lowly expressed in the midgut, fat body, silk glands and haemolymph. When day-3 fifth instar larvae were injected with LPS, the RT-PCR results showed that this gene was highly induced and it was highly expressed in the fat body and haemolymph, and the highest expression was at 6 h after injection. It is inferred that this serpin gene may play an important role in immunity of B. mori.