Clone And Sequencing Of Vitellogenin In Antheraea Pernyi And Bombyx Mori

This paper mainly contains clone and sequencing of vitellogenin of Antheraea pernyi, cloning and sequencing of vitellogenin of Bombyx mori 5'-flanking regulatory region , analysis of vitellogenin characteristic primary sequence in silk insects.Antheraea pernyi is one of economic insects with spinning and cocooning, which belong to Saturniidae in Lepidoptera. There are many research about vitellogenin of Antheraea pernyi. In this paper, several primers are designed according to cDNA sequence of vitellogenin in Antheraea pernyi.9 DNA fragments are obtained through PCR from Antheraea pernyi genomics DNA. The vitellogenin gene sequences are also obtained through cloning, sequencing and analysis.The result shows there are 6 exons and 5 introns in Vg gene of Antheraea pernyi. The exons sizes are 2258bp,205bp,982bp,879bp,184bp,829bp; and the introns sizes are 84bp,78bp,410bp,1055bp,795bp respectively. Compared with Vg gene of other insects , we find the big first intron followed by ATG 31bp in Vg of other insects absent in Vg gene of Antheraea pernyi. This is the first exceptive insect so far. The position of other intron and the sizes of exons are rather conservative. And an open reading frame of Vg gene of Antheraea pernyi encodes 1778 amino acids. This study provide some theoretic foundation for egg research about Antheraea pernyi and other insects.The DNA fragments of 5'-flanking regulatory region of vitellogenin were amplified from the Bombyx mori(951×952) genomic DNAs by using PCR. The PCR products were cloned into pMD18-T vector, then the recombined clones were identified by plasmid PCR and they were sequenced. The 5'- flanking sequence was analyzed by DNASTAR. Compared with DNA fragments of 5'-flanking regulatory region of vitellogenin in Bombyx mori(Tokai×Asahi),we find there are four place different, but the prim transcript site, TATA box and tandem repeat sequence are the same. It offered the basic experimental conditions for studying the vitellogenin 5'- flanking regulatory element and transgenic silkworm research.Seven silk insects belong to different genera in Bombyx-cidae and Saturniidae in Lepidoptera. Compared to its vitellogenin conservative primary sequemce included signal peptide ,polyserine domain, N-linked glycosylation site and so on, we can see its homology of amino acid. Phylogenetic tree of the entire amino acid sequence of Vg in seven silk insects is constructed and the results proves the conservation among evolution of silk insects vitellogenin. The homology of Philosamia Cynthia ricini and Samia Cynthia pryeri is highest and the relation of Bombyx mandarina and Saturnia japonica in two different families is far.