Quasi Black Prickly Ant Hsp90 Gene Cloning And The Level Of Mrna Expression And Its Impact On The Development Of Research

Heat Stress Proteins (HSPs) also called Heat Shock Protein(HSPs) which can be induced in living organism synthesis by some external environmental condition stimulus such as high temperature, hunger, high pressure, tissure damage.heavy metal pollution.hypoxia and metabolic toxic. HSP90may play some rules to make the body-avoiding injure or less injure. HSP90belong to one of the most widely distributed in the heat shock protein, which regulate spatial structures of many proteins.The ant Polyrhachis vicina Roger is one of the important economic insects in China, which belongs to the genus of Polyrhachis (Formicidae, Hymenoptera, Insecta. Arthropoda). As a non-toxic and harmless ant it can be used in country’s health ministry. Polyrhachis vicina Roger possesses the characteristic of caste differentiation, complex behavior, and a special experimental material for the research of development and differentiation.In this paper. Methods of RT-PCR and RACE are first used to clone the HSP90of the ant P.vicina, and then use the methods of bioinformatics and real-time quantitative PCR to some related research. The main results are offered as follows:1. The full-length cDNA of P.vicina Heat Shock Protein90(hsp90) is2325bp, the ORF is2133bp. encoding a711-amino acid protein, contains a5’-untraslated region of129bp and a3’-untraslated region of63bp. The complete P.vcina Heat Shock Protein90cDNA sequence, named Pvhsp90. were submitted to the GenBank and assigned the accession numbers:JN100104.4.2. Bioinformatics analysis the sequence of amino acids and proteins sequence. Homology analysis shows that the Pvhsp90sequence coding protein HSP protein has extremely high similarity with other species in95%; System evolution analysis shows that the first together with Apis mellifera;The protein has higher conservative, belong to ATPases super family; Primary structure analysis shows the molecular weight of hsp90protein is82009.8Da and theoretical isoelectric point (PI) is5.41respectively; the number of basic amino acids(Arg+Lys) are118. the number of acidic amino acids(Asp+Glu) are139. its chemical formula is C6289H10440N2134O25768471.Physical and chemical properties analysis shows that there are no O-glycosylation site; there are three N-glycosylationsites,29phosphorylation sites including3Serine sites,17Threonine sites and9Tyrosine sites;the HSP90protein may be hydrophilic secreted protein. The secondary structure of hsp90protein was determined by bioinformatics software, and the result displays that it is a kind of mixed protein.Three-dimensional structure analysis show that the protein sequence with the best match template is3ekoA, the best talignment region locate in the16-205amino acid residues, sequence consistency is83.684%, evalue is8.10e-86.3. Results of real-time quantitative RT-PCR show that hsp90express in every stage but different expression levels. First instar display a minimum amount expression, from second instar to pupa express quantity decline in the larva stage adult expresstion show a significantly higher. In the adult female ant’s expression is the highest after compared with male ant and work ant, this may relevant with the laying eggs function and taking off wings after laying eggs, the more investigation are needed for those specific mechanism and funcions.