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整合素是由α和β亚基组成的异源二聚体跨膜蛋白,在调控细胞增殖中起重要作用。本实验初步研究了腰带长体茧蜂Macrocentrus cingulum整合素β亚基在胚胎增殖中所起的作用。用PCR方法克隆获得了腰带长体茧蜂整合素β基因Mcitg-β(GenBank登录号为KC784942),其开放读码框全长2 526 bp,编码841个氨基酸,含有整合素基因特有的多个功能域。Mcitg-β与已报道的昆虫整合素基因具有很高的序列相似性,系统发育分析结果表明其与膜翅目的熊蜂Bombus impatiens整合素基因核苷酸序列一致性最高,达76%。定量PCR结果表明Mcitg-β在蜂卵中大量表达,在初级胚胎及二级胚胎细胞中也有表达,但在桑葚胚中没有表达。将dsMcitg-β通过体外浸泡法导入到二级胚胎或体内注射法导入到寄主亚洲玉米螟Ostrinia furnacalis幼虫体内后,可极显著抑制腰带长体茧蜂整合素基因的表达量,降低胚胎增殖能力,并减少出蜂数量。结果初步证明Mcitg-β可能对调节多胚发育的腰带长体茧蜂的胚胎增殖起重要的作用。
Integrins are heterodimeric transmembrane proteins composed of alpha and beta subunits that play an important role in the regulation of cell proliferation. In this experiment, the role of Macrocentrus cingulum integrin beta subunit in embryonic proliferation was preliminary studied. The cloned integrin β gene Mcitg-β (GenBank accession number KC784942) was cloned by PCR. The full-length open reading frame (ORF) was 5226 bp in length and encoded 841 amino acids, which contained many unique integrin genes Functional domain. Mcitg-β was highly homologous to the published insect integrin gene. Phylogenetic analysis showed that the nucleotide sequence of Mcitg-β was consistent with that of the Bombus impatiens integrin gene from Hymenoptera. Quantitative PCR results showed that Mcitg-β was abundantly expressed in bee eggs and was also expressed in primary and secondary embryonic cells but not in morula. In vitro induction of dsMcitg-β into secondary embryos or in vivo injection into the host Ostrinia furnacalis larvae could significantly inhibit the expression of integrin gene and decrease the ability of embryonic proliferation, And reduce the number of bees. The results initially showed that Mcitg-β may play an important role in embryonic proliferation that regulates polyembryonic development.