摘要
从猪胃组织中提取总RNA，采用反转录-聚合酶链反应（RT-PCR）技术，经扩增得到450bp Ghrelin基因。将其克隆到pMD18-T中。经双酶切初步鉴定后，将酶切下的目的基因进一步克隆到真核表达载体PCI中，经PCR、酶切及序列鉴定，证实插入载体PCI中的片段为含有目的基因的核苷酸序列。真核表达重组质粒PCI-Ghrelin-28aa构建成功；以构建的PCI-Chrelin-28aa质粒肌肉注射（1mg／kg）昆明小鼠（体重18-20g），观察其体重变化。结果表明：注射该基因质粒5d后，实验组小鼠平均日增重高于对照组，且差异显著（P〈0．05）。尤其是注射10-15d增重最为明显。本试验构建的PCI Chrelin-28aa基因质粒对小鼠具有明显的促生长作用．并将成为新的促猪生长的基因药物。
Total RNA was extracted from tissue of pig stomach, 450 bp fragment was obtained by RT- PCR. Then it was cloned into pMD18-T. This construction was digested with the same enzymes （Mlu I and Xho I ） and ligated to the PCI vector. The recombination plasmid was identified using PCR, digestion of internal restriction enzyme, and sequencing . The results suggested that the nucleotide sequence isolated from the recombinant plasmid PCI-Ghrelin-28aa was the same as expected. The effect of PCIGhrelin-28aa on body weight gain in mice for twenty-five days was studied. Intramuscular administration at dose of ling per 1 kg of body weight of PCI-ghrelin-28 aa to mice significantly stimulated body weight gain. For the first five days, average daily gain（ADG）was greater by the PCI-Ghrelin-28aa than by control treatment（0.93 ±0.33 vs. 0.84 ±0.29 g, respectively; P 〈 0.05）,and the tendency of increased ADG was consecutively for twenty days （ P 〈 0.05）. Conclusion： The recombinant plasmid PCI-ghrelin-28aa will be an alternative gene medicine for regulating the growth of animal.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2006年第6期667-670,共4页
Journal of Jilin Agricultural University
基金
吉林省自然科学基金项目(2004557-1)
