白菜S位点糖蛋白基因的克隆与表达分析

Cloning and Expression Analysis of SLG Gene in Brassica campestris L. ssp. chinensis(L.)Makino

以白菜[Brassica campestris L.ssp.chinensis(L.)Makino]自交不亲和系为材料,采用RT-PCR技术,用特异引物对SLG基因进行克隆,获得SLG基因cDNA序列长度为1324bp,命名为BcSLG。序列分析表明:所获得的白菜BcSLG基因cDNA序列包含一完整的编码框,编码432个氨基酸,含有12个保守的半胱氨酸残基和7个N-糖基化位点。序列比对和系统进化分析表明:BcSLG与其它植物的SLG基因氨基酸序列具有较高的同源性,与大白菜和甘蓝亲缘关系最近。荧光定量PCR分析表明:BcSLG基因在自交不亲和系的柱头中表达量最高,其次是花蕾,叶片中表达最低,在自交亲和系的柱头、花蕾和叶片中相对表达较低。

Through RT-PCR technique 1 324 bp cDNA sequence of S locus glycoprotein gene （BcSLG） was obtained from self-incompatible line with specific primers in Brassica campestris L. ssp. chinensis （L.） Makino. Sequence analysis indicated BcSLG contained one open reading frame, which encoded 432 amino acids With 12 conserved cysteine residues and 7 potential N-linked glycosylation sites. Sequence alignment and phyologenetic analysis revealed BcSLG had high similarity to SLG from other plants and had closest phylogenetic relationship to that in Brassica campestris L. and Brassica oleracea L. Real-time PCR analysis showed BcSLG was highly expressed in stigmas, much lower in buds and lowest in leaves of self-incompatible line. However, in self-compatible line BcSLG was expressed in an even lower level whether in stigmas, buds or leaves.