| 木毒蛾氨肽酶N1基因克隆与表达分析 |
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| 引用本文： | 林健聪,孙悦,刘用垄,王锦达,王荣,张飞萍. 木毒蛾氨肽酶N1基因克隆与表达分析[J]. 北京林业大学学报, 2021, 43(9): 94-100. DOI: 10.12171/j.1000-1522.20210186 |
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| 作者姓名： | 林健聪 孙悦 刘用垄 王锦达 王荣 张飞萍 |
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| 作者单位： | 1.福建农林大学林学院，生态公益林重大有害生物防控福建省高校重点实验室，福建 福州 350002 |
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| 基金项目： | 国家自然科学基金项目（31600522、31601363），福建省自然科学基金项目（2017J0106），福建农林大学林学高峰学科建设项目（61201400820） |
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| 摘 要： | [目的]氨肽酶N(APN)是昆虫中肠一类重要的Bt受体蛋白,Bt细菌产生的Cry毒素对昆虫的毒杀作用机理在学术界存在一定争议,但是普遍认为毒素与Bt受体蛋白的结合是产生毒力的必要环节.本研究通过基因克隆、生物信息学分析以及不同龄期组织中的表达对木毒蛾APN1基因进行研究,为后续研究APN基因家族、其他Bt受体蛋白及Cr...
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| 关 键 词： | 木毒蛾 Bt受体 氨肽酶N 克隆 |
| 收稿时间： | 2021-05-14 |
Cloning and expression analysis of APN1 gene from Lymantria xylina |
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| Lin Jiancong,Sun Yue,Liu Yonglong,Wang Jinda,Wang Rong,Zhang Feiping. Cloning and expression analysis of APN1 gene from Lymantria xylina[J]. Journal of Beijing Forestry University, 2021, 43(9): 94-100. DOI: 10.12171/j.1000-1522.20210186 |
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| Authors: | Lin Jiancong Sun Yue Liu Yonglong Wang Jinda Wang Rong Zhang Feiping |
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| Affiliation: | 1.Key Laboratory for Prevention and Control of Major Pest in Ecological Public Welfare Forests of Forestry College of Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China2.National Engineering Research Center of Sugarcane, Fujian Agricuture and Forestry University, Fuzhou 350002, Fujian, China |
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| Abstract: | Objective Aminopeptidase N(APN) is a class of important kind of BT receptor protein in insect midgut. The mechanism of Cry toxin produced by Bt bacteria to kill insects has been controversial in the academic research, but it is generally believed that the binding of toxin and Bt receptor protein is the necessary link to its virulence. The APN1 gene of Lymantria xylina was studied by gene cloning, biological information analysis and expression patterns, with the purpose to provid a reference for the subsequent study of APN gene family, other Bt receptor proteins and the mechanism of Cry toxin. Method APN1 was cloned by cDNA from midgut of the moth as template in a PCR system. Biological analysis and real time quantitative PCR(qRT-PCR) were performed to analyze the expression pattern of LxAPN1 gene in different ages and tissues of Lymantria xylina. Result The full-length DNA of APN1 gene was cloned from midgut of Lymantria xylina and named LxAPN1. The full-length sequence of LxAPN1 was 3 159 bp, ORF was 3 054 bp, encoding 1 017 amino acids. Sequence alignment and phylogenetic tree analysis showed that LxAPN1 and LdAPN1 were highly homologous, with signal peptide at N-terminal, zinc binding site HEXXH (X18) E and conserved region GAMENWG, and GPI binding site at the end. LxAPN1 was not expressed in egg stage, and expressed in 1?7 instar larvae, the expression of LxAPN1 decreased after larval stage; the expression of LxAPN1 in intestine was significantly higher than that in head and cuticle. Conclusion LxAPN1 was successfully cloned in the midgut of Lymantria xylina. LxAPN1 and LdAPN1 are highly homologous, and the distribution of phylogenetic tree is also very similar, which not only indicates the similarity between Lymantria xylina and Lymantria dispar, but also the similarity of APN1 function between them; The expression of LxAPN1 was the highest in the second instar larvae of Lymantria xylina. And the expression was the highest in the gut from 6 instar larvae, as an Bt receptor in the intestinal of Lymantria xylina. |
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