Cloning And Characterization Of Vg And ALF Promoters From Fenneropenaeus Chinensis

Shrimp is one of the most important aquaculture species in our country. But recent years, various diseases occur frequently which caused great economic losses to shrimp farming. Vitellogenin and anti-lipopolysaccharide factor play important roles in the process of growth and anitbacterial immunization of the shrimp. Understanding the regulation mechanism of vitellogenin and anti-lipopolysaccharide of shrimp are meanful for artificial breeding and the development of aquaculture in theory and reality.In this paper, we use Fenneropenaeus chinensiss as the research object, the main results are as follows:Part1:The promoter of Vg and the coding regions of ALF, including the upstream regulatory region, were amplified by Genome Walking method. The sequence is1100bp and2114bp, respectively. The finding by analysis is that the ALF contain two intons,365bp and319bp, respectively, and the up-stream regulatory fragment of738bp.The core promoter and cis-acting elements were found using multiple web sites. The results showed that+1scores calculated is1and "TATA box" were discovered in both genes, but the "CAAT box" and "GC box" are absent. Besides, a few cis-acting elements which may affect the activity of promoter were found among the upstream regulatory region, such as NF-κB, YY1, SP1, and so on.Part2:In order to validate the functions of Vg and ALF promoters, the recombinant fluorescence expression vectors which contain different lengths of Vg or ALF gene promoters (PVg and PALF) were transfected into SF-9cells. It turned out that pEGFP, the fluorescent expression vectors, driven by4PVg and4PALF were not performed in vivo SF-9. What’s more, the Vg genes stimulated by17β-estradiol were not expressed. Especially, a few of ALF transferred vectors stimulated by lipopolysaccharide or β-1,3-glucan could expresse the green fluorescence in SF-9cells. The results suggest that PALF is an inducible promoter.