Quasi Black Prickly Ant Dpp Gene Cloning And The Level Of Mrna Expression And Its Impact On The Development Of Research

dpp (decapentapletic) gene, belonging to transforming growth factor TGF-beta (transforming growth factor beta) superfamily. exsits in the nematodes of lower animals, which play a very important role in the growth and development of insects. For example, in the early embryonic development of Drosophila, dpp gene encoding the protein in the embryo somite differentiation process, which guide the formation of the embryonic somite interval, and participate in the differentiation of the dorsal-ventral axis; dpp as downstream genes of embryonic development gene wingless and hedghog. which determine the developmental position of the Drosophila subsidiary limbs (antennae, wings and legs). The formation of the Drosophila compound eye retina and the wings is relevent to dpp genes encoding organogenesis prime factor. The protein encoded by Thbolium castaneum and the Drosophila dpp gene respective is highly similar, at the same time; an intron within the gene is also highly conserved. In the process of embryonic development. Tcdpp is express in the back and chest subsidiary at the end of the limb. This region-specific expression is concerned with the formation of Tribolium castaneum’s subsidiary limb. The study found that with the evolution of the species and the dpp gene will be differentiation in some sequence. So dpp may have a certain valve in analysis of the evolution of species. In human growth and development process, the formation of the dpp encoding fibroin has relation to the organ formation of certain size and shape. Once the abnormal dpp’s expression, it may cause cancer or organ defects. Therefore, by studying the dpp gene, the mechanisms of organogenesis and cancer will be understood fully, which may favor therapeutic drug research in certain related diseases.Polyrhachis vicina Roger belonging to the Insecta. Hymenoptera. Formicidae. Polyrhachis. That is a typical social insect and likes gregarious life. It was designated by the Ministry of Health and determined the only food and medicine used along with the ants. The group has the typical characteristics of social insects, with the polymorphism, the grade of differentiation are the female ants, the male ants and the worker ants. The group has a highly complex nervous system, between the grades; they have the complex behavior, a clear division of labor, so it is a good experimental material to study insect development.In this experiment, the Polyrhachis as object of study, first by RT-PCR and RACE techniques to obtain dpp gene cDNA sequence, second bioinformatics technique is used to analysis of nucleotide sequence, protein sequence and the evolutionary relationship. At the same time the function prediction is also completed. Last, the real-time quantitative PCR is used to research dpp gene of Polyrhachis vicina at different developmental stages and different grades mRNA expression, the obtained experimental results are as follows:1、dpp gene of Polyrhachis vicina" Full-length cDNA sequences are1866bp, bearing a length of1644bp an open reading frame which encoded protein contains547amino acid residues. The length of the3’untranslated region of104bp and exsits a typical polyadenylation signal AATAA and sequence length is the PolyA tail of13bp. The length of the5’untranslated region of118bp. The Full-length cDNA sequences of Polyrhachis vicina was named Pvdpp, and upload it to the U.S. National Center for Biotechnology Information (NCBI) GenBank database, the gene sequence’s serial-number is JX021498.2、The results of protein sequence alignments indicate that the PvDPP protein shares an overall identity with other known DPP homologues. The encoded proteins’ molecular weight is61804.7Da. bioinformatics software analyze find that theoretical isoelectric point pI=9.79, and has a typical TGFβ domain. Analysis with the SignalP program showed that the PvDPP protein may be hydrophilic nonsecreted protein. The secondary structure of PvDPP protein was determined by bioinformatics software, and the result displays that it is a kind of mixed protein. Three-dimensional structure analysis showed that the protein sequence with the best match template is2h62B. sequence consistency is81.55%, and evalue is1.26e-50.3, The real-time quantitative RT-PCR technology research dpp gene in the Polyrhachis vicina of different grades and different developmental stages of the worm mRNA expression analysis showed that:dpp gene expression in various developmental stages of Polyrhachis; the highest expression level is in the first instar; and higher expression level is the third instar and pupa; Relatively speaking, the expression of egg and the second instar is lower, dpp gene of Polyrhachis vicina embryos have high expression levels may be involved in the formation of somite interval; The dorsal ventral axis of differentiation and determine the phylogenetic position of the subsidiary limb during embryonic development. So dpp gene is a key gene during embryonic development. In adult insects, worker ants’expression is the highest, the higher is famale ants and the lower is male ants. This may be related to the grade differentiation and worker ants" social function. This can be speculated that the dpp gene has an important role in the regulation of behavior, but the specific the mechanism and function needs further study.This study for the first time from the social insects-Polyrhachis vicina successfully cloned the full-length cDNA sequence of the dpp gene. And the sequences uploaded to GenBank; Molecular bioinformatics methods to study the evolution of the dpp gene and its Nucleotide and protein sequences. Through real-time quantitative RT-PCR technology found that mRNA expression at different developmental stages and in different grades of worms are different. These findings allow us to further understand the structure and function of the dpp gene, on this basis; we can further study the role and influence of the gene for insect development.