Clone And Analysis Of CDNA Of Vitellogenin From Actias Selene Hubner

This thesis mainly contains the clone and analysis of cDNA sequence of vitellogenin in one of economic insects with spinning and cocooning Actias selene Hubner,which belongs to Lepidoptera.There are little study related to moon silkworm Actias selene Hubner at home and abroad.Two subunits of vitellogenin in moon silkworm have been identified through SDS-PAGE and Western blotting,and also have expression difference in different sexs,tissues,times.Several primers are designed according to cDNA sequence of vitellogenin in Antheraea.pernyi,antheraea,yamamai,Bombyx.mori.Philosamia Cynthia ricini,Bombyx mandarina,Samia Cynthia pryeri,Saturnia japonica.The cDNA sequence of vitellogenin in Actias selene Hubner is obtained through RT-PCR to the total RNA and RACE amplification to 3'-end,which contains 5701 bases,an open reading frame which encodes 1774 amino acids,and the putative cleavage site RSRR for the proteases of the subtilisin family at position 364-367 where provitellogenin is cleaved into two subunits which molecular weight are 161.75KD and 40.79KD respectively based on amino acids,which are in accord with the result of SDS-PAGE if modification is considered after translation.The homology of bases between Actias seleneHubner, Saturniajaponica(AB190809),antheraeayamamai(AB055843), Antheraea pernyi(AB049631),Bombyx mandarina(AB055845),Bombyx mori(D13160),Samia Cynthia pryeri(AB190810),Philosamia Cynthia ricini(ab0555844).Homology identities are 83.1%,81.1%,81.0%,64.7%,64.6%,79.7%,79.2%respectively.Homology identity is high compared with other insects.A phylogenetic tree is established based on the vitellogenin cDNA of eight silk insects,the result proves the conservation of Vg in evolution of insect vitellogenin.This sequence has been submitted to GeneBank,the accession number is EF523567。...