乳白蚁属部分种类16S rRNA的克隆和鉴定
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密 惠 保
乳白蚁属部分种类16S rRNA的克隆和鉴定(8700字)
摘 要：大家白蚁（Coptotermes curvignathus Hollmgren）属鼻白蚁科，乳白蚁属，是一类在经济上极为重要的危险性害虫。广泛分布于东南亚等国家，是国家禁止进境检疫性有害生物。大家白蚁主要为害热带、亚热带林木和果树，是林木生产的重要威胁之一。大家白蚁主要通过原木、锯材以及木质包装进行传播，一旦传入，将给我国的森林资源、环境资源以及建筑业等造成难以估量的损失。随着我国农业生产的集约化、森林禁伐限伐政策的出台及进境原木零关税政策的实施，进境原木数量急剧增加，势必增加了大家白蚁传入的机会和风险，因此筛选适宜、准确、快速的大家白蚁鉴定及分类方法，采取合理的检疫措施已成为我国农林业生产上亟待解决的问题。
关键词：乳白蚁；16SrRNA；进出口检验检疫；克隆；PCR。
Termite species are part of the cloning and identification of 16S rRNA
Abstract : We are nose termites termite families, genera Coptotermes, is a kind of very important economic pest risk. Widely distributed in Southeast Asia and other countries, is against the quarantine pests. Termite damage all the major tropical and subtropical trees and fruit trees, is one major threat to forest production. We termites mainly through logs, sawn timber and wood packaging for distribution, if passed, will give China's forest resources, environmental resources and construction industries incalculable loss. With the intensification of agricultural production, forest logging ban and the introduction of the policy limit cutting logs entering the implementation of zero tariff policy, a sharp increase in the number of log entry, everyone is bound to increase the opportunities and risks of introduction of termites, therefore suitable for screening, accurate and rapid identification and classification of termites we take reasonable measures to quarantine of agriculture and forestry has become an urgent problem on..
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Key words: Termite; Rapid extraction; Clone; PCR
目 录
摘要 ……………………………………………………………………………… 1
关键词 ………………………………………………………………………… 1
1前言 ………………………………………………………………………… 2
2材料和方法………………………………………………………………………… 4
2.1材料 …………………………………………………………………………………4
2.1.1实验对象 …………………………………………………………………… 4
2.1.2主要试剂与仪器 ……………………………………………………………4
2.2 实验方法 …………………………………………………………………………4
2.2.1 DNA提取……………………………………………………………………4 [来源：http://think58.com]
2.2.2 PCR扩增………………………………………………………………………4
2.2.3 PCR产物纯化…………………………………………………………………5
2.2.4 DNA克隆…………………………………………………………………………5
（I）连接………………………………………………………………………5
（II）大肠杆菌DH 5α感受态细胞的制备（CaCl2）…………………………6
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（III）转化………………………………………………………………………6
（IV）碱裂解法提取质粒………………………………………………………6
（V）重组质粒的PCR鉴定……………………………………………………7
2.2.5DNA序列分析及分子系统树的构建…………………………………………7
3结果和分析…………………………………………………………………………8
3.1乳白蚁属形态鉴定特征…………………………………………………………8
3.1.1大家白蚁…………………………………………………………………8
3.1.2赛庞乳白蚁………………………………………………………………8
3.1.3台湾乳白蚁……………………………………………………………8
3.1.4格斯特乳白蚁……………………………………………………………9
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3.1.5婆罗乳白蚁……………………………………………………………9
3.1.6主要乳白蚁详图…………………………………………………………10
3.2乳白蚁16sRNA基因片段的组成及变异………………………………………16
3.3分子系统树………………………………………………………………………16
4 讨论…………………………………………………………………………^………19
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参考文献………………………………………………………………………………20
致谢…………………………………………………………………………………22 [资料来源：http://think58.com]