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2025年7月31日 星期四
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Cry1Ab敏感和抗性亚洲玉米螟氨肽酶N基因的克隆及序列差异分析
徐丽娜1,常雪艳2,何康来1,王振营2,白树雄2
1. 中国农业科学院植物保护研究所
2.
Cloning and Sequencing of Aminopeptidase N Genes from Asian Corn Borer Susceptible and Resistant to Cry1Ab Toxin
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摘要 研究亚洲玉米螟(Ostrinia furnacalis)对Cry1Ab产生抗性的分子机理,对今后制定和实施合理的抗性治理策略具有重要的意义。本研究通过PCR方法克隆和测序,鉴定了Cry1Ab敏感-抗性亚洲玉米螟幼虫中肠氨肽酶N(APN)基因家族的一个成员Ofapn3,其cDNA全序列长3 591 bp,开放阅读框包括3 045 bp,编码1 014个氨基酸。预测蛋白质分子量为115.1 kD,等电点(pI)为4.44。其推导的氨基酸序列中具有鳞翅目昆虫氨肽酶典型结构特征,即N-末端具有18个氨基酸的剪切信号肽,谷氨酸锌化氨肽酶保守结构GAMEN,锌结合位点HEXXHX18E,C-末端具有22个氨基酸的糖基磷脂酰肌醇(GPI)锚信号肽。系统分类归为第3支系。与欧洲玉米螟(Ostrinia nubilalis)的Onapn3(GenBank登录号:ABL01483)同源性为96.6%。与Cry1Ab敏感亚洲玉米螟cDNA相比,抗性品系的开放阅读框中有40个碱基发生了点突变,导致氨基酸序列中有10个氨基酸改变。其中Ser735在抗性品系中突变为Pro的现象在抗性棉铃虫APN3的氨基酸突变中也被检测到。鉴定的Cry1Ab敏感和抗性亚洲玉米螟品系的Ofapn3 cDNA序列,已提交GenBank,登录号分别是EF538427和EU137839。
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徐丽娜
常雪艳
何康来
王振营
白树雄
关键词 亚洲玉米螟氨肽酶NCry1Ab    
Abstract:It is meaningful for resistance management to study the resistance mechanism of Asian corn borer (Ostrinia furnacalis) to Cry1Ab. cDNA of aminopeptidase N (APN) from Cry1Ab toxin susceptible and resistant strains of Asian corn borer was cloned and sequenced. Ofapn3, a member of aminopeptidase N families, was identified. It contained 3 591 nucleotides with a 3 045 bp open reading frame that encoded 1 014 amino acids. Putative protein sequences included 8 and 4 potential N- and O-linked glycosylation sites and a zinc metal binding site motif of HEXXHX18E, which was typical of the active sites of the zinc-dependent metalloproteases, and a highly conserved GAMEN motif, which was to form part of the active site. The sequences had a cleavable N-terminal signal peptide with 18 amino acids and a glycosylphosphatidylinositol (GPI) anchor signal peptide with 22 amino acids in C-terminal, which were also typical characters of lepidopteran APN proteins. Sequence analysis indicated that the deduced protein sequences were most similar to Onapn3 (GenBank accession No.ABL01483) from Ostrinia nubilalis with 96.6% sequence identity. The predicted protein molecular weights and isoelectric points were 115.1 kD and 4.44, respectively. Forty nucleotide differences were observed in the open reading frames which translated into 10 amino acid differences in the putative protein sequences. The sequences reported in this paper have been deposited in the GenBank database (accession No. EF538427and EU137839 for Ofapn3 from Cry1Ab-resistant strain ACB-AbR and susceptible strain ACB-BtS).
Key wordsAsian corn borer    Aminopeptidase N    Cry1Ab
收稿日期: 2010-02-08     
通讯作者: 何康来   
引用本文:   
徐丽娜1,常雪艳2,何康来1,王振营2,白树雄2. Cry1Ab敏感和抗性亚洲玉米螟氨肽酶N基因的克隆及序列差异分析[J]. , 2011, 19(1): 164-170.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2011/V19/I1/164
 
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