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中国农学通报 ›› 2012, Vol. 28 ›› Issue (34): 85-91.

所属专题: 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

棘孢木霉T4小分子疏水蛋白基因hyb2克隆及序列分析

古丽吉米拉.米吉提 范海娟 刘志华 王娜 窦恺 黄颖 王志英   

  • 收稿日期:2012-07-03 修回日期:2012-08-19 出版日期:2012-12-05 发布日期:2012-12-05
  • 基金资助:

    国家自然科学基金面上项目;黑龙江省青年科学基金项目;黑龙江省博士后科研启动金

Cloning and Sequence Analysis of Small Molecular Hydrophobin Protein hyb2 Gene from Trichoderma asperellum T4

  • Received:2012-07-03 Revised:2012-08-19 Online:2012-12-05 Published:2012-12-05

摘要:

为了深入研究生物防治菌棘孢木霉T4菌株小分子疏水蛋白hyb2的功能,基于已知小分子疏水蛋白基因hyb2部分cDNA序列设计引物,分别以棘孢木霉T4菌株菌丝体总mRNA和基因组DNA为模板,进行PCR扩增,克隆获得hyb2的全长cDNA和DNA序列,其GenBank接受号分别为JX014433和JX185070,cDNA序列编码区长度为321 bp,编码106个氨基酸。BlastP相似性分析表明该基因与深绿木霉的Tahyd5a (ABS59366)基因同源性最高为87%,SignalP信号肽分析表明N端第16和17个氨基酸中间有一个信号肽剪切位点(AFA||AP)。Pfam蛋白家族分析表明它属于hydrophobin_2 superfamily家族。将棘孢木霉T4菌株小分子疏水蛋白基因hyb2对深绿木霉ATCC74058基因组和绿色木霉Gv29-8基因组进行同源性搜索,在2个近缘种的基因组上分别获得9个和8个同源序列。其中,深绿木霉ATCC74058的hyb-a-7和绿色木霉Gv29-8的hyb-v-5疏水蛋白与棘孢木霉Hyb2相似性最高分别为87%和70%,2个序列分别位于深绿木霉ATCC74058染色体骨架5和绿色木霉Gv29-8染色体骨架22上。

关键词: 产量性状, 产量性状

Abstract:

In order to investigate the function of Small molecular hydrophobin protein hyb2 in biocontrol fugi Trichoderma asperellum T4, the series of primers were designed based on the known cDNA fragments of small molecular hydrophobin protein hyb2, with the total RNA and the genome DNA extracted from Trichoderma asperellum T4 as the template, using PCR method, the full length cDNA and DNA sequence of hyb2 gene finally were obtained, their accession numbers were JX014433, JX185070, respectively. The cDNA sequence of hyb2 was 321 bp in length, encoding 106 amino acids. BlastP search indicated that hyb2 shared the highest similarity 87% with Tahyd5a of T. atroviride (ABS59366). Furthermore, signalP prediction showed that the hyb2 amino acid sequence was cleaved by signal peptidase between positions 16 and 17 (AFA||AP). Pfam protein family prediction showed that Hyb2 protein belonged to hydrophobin_2 superfamily. To obtain homologous gene sequences of hydrophobin protein hyb2 from the genome databases of T. virens Gv29-8 and T. atroviride ATCC74058, the amino acids sequences of hyb2 from T. asperellum T4 was used to as queries, resulting that 9 and 8 hydrophobin sequences with high similarity were obtained, respectively. Hydrophobin proteins hyb2 shared the highest 87% and 70% similarity with hyb-a-1 and hyb-v-2, located on T. atroviride ATCC74058 scaffold 5 and T. virens Gv29-8 scaffold 22, respectively.

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