传疟媒介大劣按蚊和斯氏按蚊基因组dna遗传多态性研究病原生物学专业论文.docxVIP

安徽理工大学硕士学位论文Abstract 安徽理工大学硕士学位论文 Abstract Objeefive To analyze the genomic DNA polymorphism in An．6矗rus and An．stephensi of different susceptible to Plasmdium yoelii，SO as to explore host genotype by parasite genotype interactions underlying the susceptibility of anopheline mosquitoes to plasmodium． Methods The genomie DNA was extracted from the adults of An．dims and An．stephensi．On the one hand，the genomie DNA for both species were amplified by RAPD，and then the amplified products were carried on agaxose gel electrophoresis．The gel-purified RAPD bands which had similar mobility WCl-e isolated and sequenced．Sequence alignment was performed by using DNASIS program．On the other hand,the rDNA ITS2 region for both species we糟amplified by PCR and sequenced,and the amplified products were analyzed by restriction endonuclease digestion(RFLP)．The correspondent sequence of An．西h口and An．stephensi were searched from GenBank．All these sequences we∞aligned and compared using Clustal W program,and the phylogenetie tree was constructed by MEGAprogram． Results The genomic DNA of An．dirua and An．stephensi were successfully amphfied by RAPD，and there was obvious difference in their RAPD fingerprinting on genomie DNA,respectively．Sequence alignment of four pairs of RAPD bands which had similar mobility between different anopheline mosquitoes showed that each pair of RAPD bands WOIe differed in GC content and Simple Sequence Repeat,and lheir difference Wele in the arrange from 60．0％to 61．9％in nuclentide sequences．It was demonstrated that there were poly】morphism in the length and construction of RAPD bands which had similar mobility．Moreover,there撇significantly different from the RFLP patterns ofrDNA ITS2 in An．dims and An．stephensi by restriction endonuelease Xho L the length of ITS2 region was 715bp in An．舭and 466bp in An．stephensi, Sequence alignment of ITS2 region in An．dirus and An．stephensi showed that ITs2 region WCle differed in GC content and Simple Sequence Repeal，an