tailieunhanh - Insecticides Basic and Other Applications Part 3

Tham khảo tài liệu 'insecticides basic and other applications part 3', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | The Pyrethroid Knockdown Resistance 29 Kdr site J I . . Sample DNA WWWWWWWW ATGTC ACTTC WWWWWW wvw vww G A WW Primers Twwv Alelie specific amplification wwwwwwww 70bp wwwwwwwwwww 90bP Amplification wwwwwwwwwwwwa WvVv G WQ Specific probes jWAAAMT Fig. 4. Examples for kdr genotyping based on PCR methods. A - Allelic specific PCR with specific primers in different orientations B - Allelic specific PCR with specific primers in the same orientation but with additional and differently sized GC n tails in addition to a mismatch in the 3rd base before the 3 -end C - TaqMan assay based on specific probes with a different luminescence for each allele. Figure adapted from Yanola et al. 2011 . 30 Insecticides - Basic and Other Applications al. 2008 . However comparison between HOLA and pyrosequencing revealed more specificity for this latter method in the diagnostic of the kdr mutation Leu1014Phe in Cx. quinquefasciatus Wondji et al. 2008 . Sequencing of regions that encompass the SNP allows a direct visualization of the nucleotide allele sequences eliminating the problem of unspecific amplification or hybridization of PCR based protocols. Moreover it enables visualizing potential novel variations that would never be identified by PCR diagnostic SNP techniques. However sequencing in large scale is much more expensive than the aforementioned genotyping tools. It is also mandatory that the eletropherograms generated have a clean profile so that the heterozygous individuals can be undoubtedly discriminated. 7. Conclusions New strategies for arthropod control based on the release of laboratory manipulated insects that would suppress or substitute natural populations are being tested in the field with great prospect. The release of transgenic insects carrying a dominant lethal gene RIDL Black et al. 2011 or of mosquitoes with the intracellular Wolbachia that lead to refractoriness to other parasites Werren et al. 2008 are currently the most discussed .