[Construction of suppression subtracted cDNA library of deltamethrin-resistant Aedes albopictus].
作者信息
Wu Jia-hong, Zhao Tong-yan, Dong Yan-de
机构信息
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing, China.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Aug;24(4):281-4.
OBJECTIVE
To construct the suppression subtracted cDNA library of deltamethrin-resistant Aedes albopictus.
METHODS
Total RNA was extracted from the deltamethrin-resistant (R-lab) and -sensitive (S-lab) isolates, mRNA was obtained after purification. Double stranded cDNAs were synthesized after reverse transcription. Two subtractions were performed by suppression subtractive hybridization with S-lab as tester and R-lab as driver or S-lab as driver and R-lab as tester. Enriched different expressed cDNA was cloned into pMD18-T vector to construct subtractive libraries.
RESULTS
The subtracted cDNA libraries contained 580 and 477 positive clones respectively. The PCR results of 150 clones picked randomly from each library showed that the positive ratio of constructed cDNA libraries was 93%, with a length of cDNA fragments ranged from 150bp to 750bp.
CONCLUSION
The suppression subtracted cDNA library of deltamethrin-resistant Ae. albopictus is constructed.
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