杂种和纯种猪之间脂肪组织差异表达基因的分离,克窿和序列分析.pdfVIP

业生物技术学报 Journal of Agricultural Biotechnology 2005,13 (5): 60 1~607 ·研究论文· Isolation, Cloning and Sequence Analysis of Differentially Expressed Genes in the Backfat Tissue between Hybrids and Parents in Pigs* REN Zhu-Qing XIONG Yuan-Zhu** DENG Chang-Yang LEI Ming-Gang ZUO Bo ZHENG Rong LI Feng-E JIANG Si-Wen JIANG Xun-Ping 邾 In order to reveal molecular basis of heterosis, Large White (LW), an introduced European pig breed, and Meishan (MS), a Chinese indigenous pig breed, were selected to hybridize directly and reciprocally in the present experiment. mRNA differential display (DD) technique was performed to identify genes that were differentially expressed among backfat tissues of hybrids (LW伊MS, MS 伊LW) and purebred (LW伊LW, MS 伊MS) pigs. The 10 anchor primers in combination with ten arbitrary primers (100 sets in total) were used and nearly 1 500 reproducible bands were observed in polyacrylamide gel. The 40 differentially displayed bands were selected to clone and sequence. The 36 out of 40 ESTs were proved to be novel and the sequences were submitted to GenBank (accession No. CV507051~CV507087), and the other 4 showed similarity to known genes published in GenBank. Three among 36 novel ESTs were chosen for further identification with semi-quantitative RT-PCR. The result showed that 2 ESTs were differentially expressed, and the 3rd showed no obvious difference between hybrids and purebreds. In order to reduce the percentage of false positive of DD, RNA pools of four types of pigs were constructed respectively by mixing 6 samples from 6 pigs of the sa