反义核酸抑制抗敌百虫淡色库蚊扩增酯酶mRNA体外翻译的研究

中国寄生虫学与寄生虫病杂志 ›› 2001, Vol. 19 ›› Issue (6): 4-335.

反义核酸抑制抗敌百虫淡色库蚊扩增酯酶mRNA体外翻译的研究

朱淮民¹;邹亚丁²

第二军医大学病原生物学教研室上海200433 (朱淮民);贵州省疾病预防控制中心地方病防治所贵阳550001(邹亚丁)

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-12-30 发布日期:2001-12-30

Inhibition of in vitro Translation of Esterase mRNA of Dipterex-Resistant Mosquito (Culex pipiens pallens) by Antisense Nucleic Acids

ZHU Huai min ¹;ZOU Ya ding ²

1 Department of Etiologic Biology;Second Military Medical University;Shanghai 200433; 2 Institute of EndemicDiseases;Guizhou Provincial Center for Disease Prevention and Control;Guiyang 550001

Received:1900-01-01 Revised:1900-01-01 Online:2001-12-30 Published:2001-12-30

摘要/Abstract

摘要： 　　目的　用反义核酸抑制抗敌百虫淡色库蚊扩增酯酶mRNA体外翻译。　方法　人工合成互补于抗性库蚊酯酶mRNA翻译起始点 18碱基 ,与淡色库蚊mRNA退火后加入无细胞翻译体系 ,进行体外翻译 ,翻译产物用SDS 聚丙烯酰胺凝胶电泳进行分析。　结果　 6 μmol/LODNs可抑制 5 0 %特异性酯酶翻译量 ,2 0 μmol/LODNs抑制 80 %特异性酯酶翻译量。电泳结果显示条带浓度接近敏感蚊虫所表达的酯酶量。　结论　针对抗敌百虫淡色库蚊扩增酯酶mRNA翻译起始点的反义核酸在体外能有效地抑制其mRNA的翻译

关键词: 有机磷抗性, 酯酶, 反义核酸, 淡色库蚊

Abstract: 　Objective To examine the inhibitory effect of antisense nucleic acid on the in vitro translation of esterase mRNA from dipterex resistant Culex pipiens pallens . Methods 18 mer nucleic acid was synthesized and complementary to the translation initiation site of mRNA of dipterex resistant mosquitoes. The ODNs were annealed to the corresponding mRNA molecules and they were added to rabbit reticulocyte cell free system. The translation products were analyzed by SDS PAGE. After fixing, the gel was exposed to X ray film by autoradiography for analysis of protein synthesis. Results Six μmol/L of ODNs elicited a 50% reduction in specific protein expression , and 20 μmol/L of ODNs inhibited the expression of esterase by 80%. The SDS PAGE showed that the band of reduced amounts of 65 kDa protein for resistant mosquito was almost the same as that for sensitive sample. Conclusion Antisense oligonucleic acids to the esterase mRNA of dipterex resistant mosquito could effectively inhibit its in vitro translation.

Key words: organophosphorus resistance, esterase, antisense nucleic acid, Culex pipiens pallens

朱淮民;邹亚丁. 反义核酸抑制抗敌百虫淡色库蚊扩增酯酶mRNA体外翻译的研究[J]. 中国寄生虫学与寄生虫病杂志, 2001, 19(6): 4-335.

ZHUHuaimin;ZOUYading. Inhibition of in vitro Translation of Esterase mRNA of Dipterex-Resistant Mosquito (Culex pipiens pallens) by Antisense Nucleic Acids[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2001, 19(6): 4-335.

[1]	吕文祥, 程鹏, 彭荟, 王海洋, 刘宏美, 王海防, 郭秀霞, 公茂庆, 刘丽娟. 淡色库蚊不同发育阶段肠道菌群多样性分析[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(4): 460-467.
[2]	周洁, 李春晓, 兰策介, 高剑, 刘钦梅, 孙爱娟, 赵彤言. 淡色库蚊与骚扰库蚊触角嗅觉基因表达分析[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(4): 453-457.
[3]	吴松泉1 *，王光丽1，周武1，钟石根2. 淡色库蚊转铁蛋白在毕赤酵母中的分泌表达及其抑菌活性的初步研究[J]. 中国寄生虫学与寄生虫病杂志, 2014, 32(1): 8-38-41.
[4]	张玉勤, 刘吉起, 郭祥树, 唐振强, 赵旭东, 周瑞敏, 赵奇. 河南省淮滨县中华按蚊的抗药性研究[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(6): 18-493-495.
[5]	张浩1, 2, 张威2, 朱燃2, 诸葛洪祥1 *. 印度殖盘吸虫神经系统的乙酰胆碱酯酶定位观察[J]. 中国寄生虫学与寄生虫病杂志, 2011, 29(1): 17-71-73.
[6]	张玲敏;孙家梅;吕慧芳;施海瑛. α-三噻吩对白纹伊蚊幼虫蛋白质、酯酶及脂质过氧化的影响[J]. 中国寄生虫学与寄生虫病杂志, 2010, 28(1): 11-49.
[7]	李士根. 淡色库蚊抗敌敌畏和抗氯氰菊酯品系的抗性演变[J]. 中国寄生虫学与寄生虫病杂志, 2009, 27(4): 18-312.
[8]	宋锋林;赵彤言;董言德;陆宝麟. 淡色库蚊氯菊酯抗性品系和敏感品系的生物学特性[J]. 中国寄生虫学与寄生虫病杂志, 2005, 23(1): 10-42.
[9]	王新国;甄天民;谭文彬;王怀位;公茂庆;孙传红;赵玉强. 淡色库蚊DDVP抗性与酯酶活性的相关性[J]. 中国寄生虫学与寄生虫病杂志, 2003, 21(2): 8-92.
[10]	王志钢;战志胜;刘宏超;海秀平;尚文旭;朱建新. 淡色库蚊冬季爆发原因及防制措施[J]. 中国寄生虫学与寄生虫病杂志, 2003, 21(1): 26-61.
[11]	吴明玮;张玲敏. 白纹伊蚊乙酰胆碱酯酶基因片段序列分析[J]. 中国寄生虫学与寄生虫病杂志, 2002, 20(3): 18-187.
[12]	李焱;余新炳;吴忠道;李晖婷;包俊英;邵筱. 序列标签法寻找日本血吸虫新基因[J]. 中国寄生虫学与寄生虫病杂志, 2002, 20(1): 9-34.
[13]	耿志辉;方艳秋;刘利;石毅;李淑红. 弓形虫病大鼠T淋巴细胞和脾微量元素的测定[J]. 中国寄生虫学与寄生虫病杂志, 2001, 19(6): 11-359.
[14]	张恩英;王仁赉. 三带喙库蚊和淡色库蚊胸肌线粒体琥珀酸脱氢酶比活力测定[J]. 中国寄生虫学与寄生虫病杂志, 2001, 19(5): 20-320.
[15]	田海生;朱昌亮;高晓红;马磊;沈波;李秀兰;吴观陵. 淡色库蚊对溴氰菊酯抗药性和敏感性相关基因克隆和序列分析[J]. 中国寄生虫学与寄生虫病杂志, 2001, 19(4): 1-197.