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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2016, Vol. 36 Issue (4): 63-68    DOI: 10.13523/j.cb.20160410
研究报告     
RNA干扰沉默KAT基因对蜜蜂合成10-HDA的影响
刘丽, 杨晓慧, 王瑞明
齐鲁工业大学 生物工程学院 济南 250353
The Effect of KAT Gene Silencing by RNAi on the Synthesis of 10-HDA in Bees
LIU Li, YANG Xiao-hui, WANG Rui-ming
Department of Biology Engineering, QILU University of Technology, Jinan 250353, China
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摘要：

合成KAT基因的双链RNA,在蜜蜂体内进行RNA干扰,利用半定量RT-PCR技术和高效液相色谱法,分别检测RNA干扰后KAT基因的表达量和蜜蜂10-HDA的合成量,初步探讨沉默KAT基因与蜜蜂合成10-HDA的关系。结果显示,在蜜蜂体内注射2μl的dsKAT其成活率最高,且注射5天后,与空白组相比,KAT基因的表达量最低,蜜蜂头部10-HDA的合成量降低效果显著(P<0.05),蜜蜂上颚腺表面腺体相对干瘪、疏松。成功实现了蜜蜂体内KAT基因的沉默,且初步推测出KAT基因参与10-HDA的合成途径,阐明了KAT基因与蜜蜂合成10-HDA的关系。这为利用RNA干扰技术分析其他蜜蜂基因功能提供了借鉴,为进一步完善10-HDA的生物合成途径奠定了基础,为通过提高基因表达量来提高10-HDA的产量提供了依据。
关键词： RT-PCR;  10-HDA;  RNA干扰;  KAT    
Abstract:

The double-stranded RNA of KAT gene was synthesized for RNA interference (RNAi)in bees. After RNAi, the expression of KAT gene and the synthesis of 10-HDA were detected respectively by semi-quantitative RT-PCR and HPLC in order to investigate the relationship between the silencing of KAT gene and the amount of 10-HDA. These results showed that the bees which were injected with 2μl dsKAT had the highest survival rate. And when the bees were injected for five days,the KAT mRNA levels seemed the lowest and the secretion of 10-HDA in honeybee workers showed a significant decrease(P<0.05)relative to the control group which was injected with DEPC water.And after knockdown of KAT, the glands of the bees MGs appeared to be wizened and loose .The KAT gene in bees was silenced successfully,which speculated that KAT might be involved in the biosynthetic pathway of 10-HDA and described the relationship between KAT gene and the synthesis of 10-HDA. It provides a reference for analysing the function of other genes in honeybees by RNAi, and has laid a foundation for the further perfection of 10-HDA biosynthetic pathway. In addition, it provides the basis for improving the yield of 10-HDA by increasing the expression of genes.
Key words: 10-HAD    RNAi    KAT    RT-PCR
收稿日期: 2015-12-31 出版日期: 2016-02-15
ZTFLH:  Q789  
基金资助:

国家自然科学基金(31201281,31171727)、山东省优秀中青年科学家科研奖励基金计划(BS2013SW029)资助项目
通讯作者: 王瑞明     E-mail: ruiming3k@163.com
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刘丽
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引用本文:

刘丽, 杨晓慧, 王瑞明. RNA干扰沉默KAT基因对蜜蜂合成10-HDA的影响[J]. 中国生物工程杂志, 2016, 36(4): 63-68.

LIU Li, YANG Xiao-hui, WANG Rui-ming. The Effect of KAT Gene Silencing by RNAi on the Synthesis of 10-HDA in Bees. China Biotechnology, 2016, 36(4): 63-68.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160410    或    https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I4/63

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