Study On The Influence Mechanism Of Environmental Factor On Drosophila Population Differentiation

To further explore the impact of environment(food)on the reproductive selection of Drosophila,that is,to verify the effect of partial deletion of the ejaculation regulatory region of the Drosophila Est-6 esterase gene found in the previous work of this group on the mating behavior of Drosophila,in this experiment,Drosophila melanogaster was used as the research object,and the adeno-associated virus vector PAAV-ALSTR-IRES-EGFP was recombined using SLIC seamless cloning technology to construct the PAAV-ESTed6-IRES-EGFP plasmid.The green fluorescent protein and Est-6 esterase in the system are under the natural promoter containing the deletion of the EDRR part of Drosophila.Then the plasmid was injected into the embryo to construct an ESTed transgenic fruit fly with the help of microinjection,followed by fluorescence detection? PCR verification and one-generation sequencing proved that the transgenic fruit fiy was successfully constructed.Then we analyzed the mating selectivity between the CMA offspring Drosophila population and the offspring ESTed transgenic Drosophila as well as the difference in expression of EST-6 enzyme activity.It is proved that partial deletion of the EDRR regulatory region of Drosophila EST-6 gene will increase the expression of EST-6,and further improve the reproductive selectivity of Drosophila,which will lead to the reproductive isolation of Drosophila.The research results show that:(1)Using the new generation of Seamless Cloning/In-Fusion Cloning,the Alst R gene and its promoter synapsin on the original plasmid PAAV-ALSTR-IRES-EGFP were replaced with the EST-6 gene and Ejaculatory duct regulatory region of the 18th generation CMY fruit fly discovered in the previous work.At the same time,after one generation of gene sequencing and sequence comparison,the recombinant plasmid was successfully constructed.(2)The fluorescent detection of the reporter gene enhanced green fluorescent protein EGFP on the recombinant plasmid proved that the plasmid was successfully transferred and expressed,and the ESTed transgenic fruit fly was successfully constructed.(3)The results of in vitro enzyme activity measurements indicate that Est-6 activity in the head of the transgenic fruit fly population is much higher than that of the CMA fruit fly population.(4)Through mating experiments between the CMA offspring Drosophila population and the offspring ESTed transgenic Drosophila population,we found that there were clearly visible differences in reproductive selectivity.