ZHAO Hong-kun,LI Qi-yun,WANG Yu-min,et al.Isolation and Analysis of Soybean Kunitz Trypsin Inhibitor Gene from Glycine tomentella[J].Soybean Science,2010,29(02):191-194.[doi:10.11861/j.issn.1000-9841.2010.02.0191]
多年生野生大豆Kunitz型胰蛋白酶抑制剂基因的克隆及分析
- Title:
- Isolation and Analysis of Soybean Kunitz Trypsin Inhibitor Gene from Glycine tomentella
- 文章编号:
- 1000-9841（2010）02-0191-04
- 关键词:
- 多年生野生大豆; Kunitz型胰蛋白酶抑制剂; 原核表达
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 利用RT-PCR方法，从对蚜虫高抗的多年生野生大豆短绒野大豆（G. tomentella）(2n=78)未成熟子叶中扩增到了大豆Kunitz型胰蛋白酶抑制剂同源基因单一目的片段，该基因和蛋白分别被命名为KTiPW54和KTiPW54。序列分析表明：该片段为654 bp的完整编码序列，编码218个氨基酸残基，编码的蛋白与栽培大豆(G.max)、野生大豆(G.soja)、短绒野大豆(G. tomentella)的Kunitz型胰蛋白酶抑制剂基因编码蛋白高度保守的区域一致。将KTiPW54编码区克隆到表达载体pTWIN1，在宿主菌BL21（DE3）中经IPTG诱导获得高效表达。
- Abstract:
- A single soybean Kunitz trypsin inhibitor gene,KTiPW54, was isolated from immature cotyledons of perennial wild soybean (G. tomentella) with high aphids resistance by RT-PCR. Sequencing analyses revealed a cDNA sequence of 654 bp long and encoded 218 amino acids. The deduced amino acid sequence of KTiPW54 exhibited high similarities to those of soybean Kunitz trypsin inhibitor. The coding region was fused to the intein of pWIN1 and over-expressed in E.coli BL21(DE3) induced by IPTG.
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备注/Memo
基金项目：国家转基因专项资助项目（2008ZX08004-004）；农业部生物资源保护与利用资助项目。