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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321bp,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。
In order to further study the function of hyb2, a small molecule hydrophobin of the strain of Bacillus thuringiensis T4, the primers were designed based on the partial cDNA sequence of the hyb2 known small-molecule hydrophobin gene. The total mRNA and genomic DNA as a template for PCR amplification. The full-length cDNA and DNA sequence of hyb2 were cloned. The GenBank accession numbers were JX014433 and JX185070, respectively. The coding region of the cDNA sequence was 321 bp and encoded a protein of 106 amino acids. BlastP similarity analysis showed that the gene shared 87% homology with Tahyd5a (ABS59366) gene of T.cerevisiae. SignalP signal peptide analysis showed that there was a signal peptide cleavage site (AFA) in the middle of the 16th and 17th amino acids || AP). Analysis of the Pfam family of proteins indicates that it belongs to the hydrophobin_2 superfamily family. The hyb2 small-molecule hydrophobin gene hyb2 was used to search the genome of T. denticulatii ATCC74058 and Trichoderma viride Gv29-8 for homology search. Nine and eight Homologous sequence. Among them, hyb-a-7 of Trichoderma viride ATCC74058 and hyb-v-5 hydrophobin of Trichoderma viride Gv29-8 had the highest similarity of 87% and 70% to Hyb2, respectively. The two sequences were located at Trichoderma viride ATCC74058 chromosomal framework 5 and Trichoderma viride Gv29-8 chromosomal framework 22.