猪SOCS3基因克隆及其在不同生长阶段表达差异研究

从杜长大母猪的肠系膜脂肪中提取基因组RNA，用RT-PCR扩增SOCS3基因，获得1条约332 bp的片段，以pGEM-T Easy vector 为载体，将该基因片段克隆到大肠杆菌(Escherichia coli)DH5α中。从筛选的阳性克隆中分离出SOCS3基因，测定其序列，分析表明，该片段为 SOCS3 cDNA的部分序列，与GenBank中登录的猪SOCS3 cDNA部分核苷酸序列同源性达到100%。以SOCS3基因片段的克隆为基础，构建了优化的半定量RT-PCR法，以18s rRNA为内标，研究不同生长阶段猪肠系膜脂肪中SOCS3基因表达的差异。结果发现，从初生到90 kg，SOCS3基因在肠系膜脂肪中表达量呈逐渐增加的趋势，其中90 kg较初生时增加了141％（p<0.05）。

Cloning and the Difference Expression of Porcine SOCS3 Gene at Different Stages

Genome RNA was extracted from mesentery adipose of porcine (Duroc×Landrace×Yorkshire) and suppressor of cytokine signaling -3(SOCS3) mRNA was amplified using RT-PCR. A DNA fragment about 332 bp in length was obtained and the PCR product was cloned into pGEM-T vector. SOCS3 gene was isolated and sequenced from the positive clones screened. Sequence analysis suggested that this fragment corresponded to a portion of the sequence of SOCS3 cDNA, its homology with SOCS3 from GenBank was up to 100%. Based on the SOCS3 gene clone, an optimal semi-quantitative RT-PCR method was successfully constructed. Using 18s rRNA as inner control, the difference of SOCS3 gene expression at different stages of swine was studied. The results showed that the amount of SOCS3 gene expression increased from 1 kg to 90 kg. and the SOCS3 mRNA level at 90 kg significantly increased by 141% (p<0.05) as compared with 1 kg.