马铃薯小热激蛋白基因sHSP-F的克隆及植物表达载体的构建

发布时间：2018-06-16 22:24

  本文选题：马铃薯 + 小热激蛋白　； 参考：《分子植物育种》2017年04期

【摘要】：热激蛋白(heat shock proteins,HSP)是生物体在不利环境条件因素刺激下应激合成的一组在进化上高度保守的蛋白质。前期转录组测序的结果发现马铃薯Favorita的小热激蛋白(small heat shock proteins,s HSPs)基因(PGSC0003DMG400009255)在接种晚疫病菌(Phytophthora infestans)24 h后表达量显著上调。因此,以马铃薯Favorita为材料,根据PGSC0003DMG400009255基因序列设计引物,并在引物5'端加上Bam HⅠ和SalⅠ酶切位点,从接种P.infestans 24小时后的Favorita的RNA中通过RT-PCR的方法获得PGSC0003DMG400009255的基因片段,并命名为s HSP-F,该基因最大开放阅读框(ORF)为594 bp,编码197个氨基酸。通过酶切连接将s HSP-F连接至表达载体p CAMBIA1301中。通过测序和酶切验证,表明s HSP-F基因成功克隆到表达载体中,该工作为进一步研究该基因的功能提供了基础。
[Abstract]:Heat shock protein (shock) is a group of highly evolutionarily conserved proteins stimulated by adverse environmental conditions. The results of transcriptome sequencing showed that the expression of small heat shock protein (HSPs) gene PGSC0003DMG40000009255 in potato Favorita was significantly up-regulated after inoculation with Phytophthora infestans)24. Therefore, based on the sequence of PGSC0003DMG400009255, the primers were designed from Favorita of potato, and the fragment of PGSC0003DMG400009255 was obtained by RT-PCR from Favorita RNA inoculated with P.infestans 24 hours after inoculation with Bam H 鈪，

本文编号：2028310