Gene Related To Wool Fineness Screening With DDRT-PCR

Wool production has an important economic value.Wool fiber diameter is an important assessment of the wool quality.in the same species,the same age,the same feeding and the growth environment,genetic differences between individuals on the growth of wool play a decisive role.Xinjiang is an important area of wool production,XinJi fine-wool sheep is the Chinese major variety of high quality wool production.184 two-year-old of XinJiang superfine wool sheep (XinJi fine-wool sheep) fibre diameter were detected,the study select the fiber diameter of the very crude and the very fine of the six fine-wool sheep,take their skin composing the RNA pool as an experimental sample,the application of mRNA differential display reverse transcription PCR technique to separate differential expressed genes in differential fiber fineness of the sheep shoulder skin,with a view to early selection for fine-wool sheep to provide theoretical and technical reference.The experiment choose three anchor primers,eight random primers,the 24 primer pairs be composed to DDRT-PCR.PCR products were analyzed by polyacrylamide gel electrophoresis,and the differential displayed genes were reclaimed,separating 53 differential expressed genes as templates of second PCR.The purified genes were cloned by pGM-T vector and verified by reverse Northern blot analysis,screening out false-positive fragments,13 positive fragments will be sequenced.In 13 sequenced fragments,there are three fragments not found primer sequences,two fragments are inconsistent with their length when cutting from polyacrylamide gel.The sequences of the remaining eight fragments are analysed in GenBank,including six unknown function genes,two putative function genes.One similar to LaminB gene(LMNB),and another sequence blast reveal that it homology to SHROOM2 genes and GPR143 gene.At the same time in this experiment,the gene expression of Bâ…¢B4 in Xinji fine-wool sheep skin was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method,The relationship between relative gene expression of Bâ…¢B4 and fineness wool were analyzed by using 18SrRNA as internal standard.The ratio of integrated optical density (IOD) of Bâ…¢B4 PCR and 18SrRNA PCR were analyzed by Independent Samples T Test.The result indicated how gene expression of Bâ…¢B4 influence fineness of wool is negative significant correlation.