Analysis Of Soluble Protein And Genomic Polymorphic DNA Of Three Ordinary Species Of Cockroaches In Anhui Province

Objective To find scientific classification methods for cockroach analysis of discrepancies among three ordinary species of cockroaches in Anhui province has been made by comparing the soluble protein strips and the genomic DNA polymorphic . The soluble protein strips and the genomic DNA fingerprints of three ordinary species of cockroaches in Anhui province were gained by using SDS-PAGE and RAPD technique. Methods The specimens of cockroaches were collected in Anhui province , and were identified by morphology . Firstly the soluble protein liquid were made by the way of freeze and melt Homogenate. SDS-PAGE was carried out in the liquid. Dyed by Coomassie brilliant blue G250 , the molecular weight of main protein strips of the cockroaches were measured and calculated by standard curve . Secondly the Genomic DNA of the three ordinary species of cockroaches was extracted, and identified and quantitative analysed .The total volume of reaction and conditions of RAPD was defined . Five primers with different arbitrary single-chain polynucleotide sequences were selected. The groups of DNA were amplified by RAPD , and the DNA fingerprint maps were gained by running agarose gel electrophoresis . the genetic distances among three ordinary species of 6 cockroaches were calculated . Lately the soluble protein strips and the genomic DNA polymorphic of three ordinary species of Cockroaches were studied and analysed Results The SDS-PAGE experiment suggested there exist some discrepancies More than 20 strips were identical on the SDS-PAGE gel the numbers of female and male of P.f. are 31 strips and 29strips ; the numbers of female and male of P.am. are 30strips and 28strips; the numbers of female and male of B.g. are 28strips and 25strips . the numbers and locations of protein bands were different among these species . It was also obvious that the positions of the main protein bands in B.g. were different from those in two Periplaneta species , and the positions of the main protein bands in female insect were different from those in male insect Little difference was founded in the bands of two Perz~planeta species with only a slightly different appearance in their densities . The RAPD experiment suggested there exist some discrepancies in the amplified DNA fragments of three species. The DNA maps that the numbers and size of DNA bands were different among the species have gained by RAPD . The common fragments indicated some degree homogeneity of three species genomes , their distinctive fragments and intensity suggested the diversity among the species . The sizes of bands in P.am. that were more intensive than others obtained by primer OPA-8 were 0.73kb and 0.40kb , the bands in P.f. were 0.70kb , the bands in B.g. were 0.52kb. The sizes of bands in P.am. that were more intensive than others obtained by primer OPA-1O were 0.33kb , the bands in P.f. were 0.80kb , the bands in B.g. were 0.33kb . The sizes of bands in P.am. that were more intensive than others obtained by primer OPA-15 were 0.50kb and 0.70kb , the bands in P.f. were 0.70kb and 0.85kb , the bands in B.g. were 0.32kb. 0.53kb. 0.61kb. 0.85kb. 1.00kb. 1.10kb and 1.20kb . The sizes of bands in P.am. that were more intensive than others obtained by primer OPA-16 were 0.30kb.