摘要
目的探讨PC-1基因高表达与肥胖、胰岛素抵抗及2型糖尿病的关系。方法构建pcDNA3.1/PC-1转基因质粒,经显微注射的方法获得G0代转基因小鼠;用PCR、Southern印迹、RT-PCR和Western印迹方法检测PC-1转基因小鼠的基因组整合和组织表达;对转基因小鼠进行腹腔葡萄糖耐量试验(IPGTT)和胰岛素耐量试验(ITT)。结果测序证明克隆的PC-1cDNA序列与GenBank相应序列相比无误,转基因质粒中PC-1插入方向正确;鼠尾基因组DNA PCR和Southern印迹表明共获得8个PC-1转基因阳性的G0代小鼠,分别与野生型C57BL/6J小鼠交配后获得5个F1代PC-1转基因小鼠系;RT-PCR、Western印迹证实该基因在2个单系的骨骼肌和肝组织中有稳定表达;与野生型小鼠相比,转基因阳性小鼠的体重、空腹血糖、IPGTT和ITT结果无明显变化。结论成功构建稳定表达PC-1基因的转基因小鼠系,研究结果表明该基因在相关组织中的高表达不足以触发肥胖、胰岛素抵抗或2型糖尿病的发生。
Objective To explore the association of high expression of PC-1 gene with obesity, insulin resistance and type 2 diabetes. Methods The recombined pcDNA3.1/PC-1 vector was constructed and a model of Go transgenie mice was established by mieroinjeetion. The integration of the foreign gene was tested by PCR and Southern blotting and its expression was examined by RT-PCR and Western blotting. Intropefitoneal glucose tolerance test （IPGTT） and insulin tolerance test （ITT） were performed to study the function of PC-1 gene in transgenic mice. Results The sequence of the recombined PC-1 gene was confirmed by direct sequencing. PCR and Southern blotting results showed that 8 of the total GO transgenic mice were positive with PC-1 gene. These mice were mated with C57BL/6J wildtype mice and 5 lines of PC-1 transgenic mice were established. PC-1 gene was found to be stably expressed in skeletal muscle and liver in 2 lines by RT-PCR and Western blotting. As compared with wildtype mice, significant changes in body weight, fasting blood glucose, IPGTT and ITT were not found. Conclusion A transgenic mice model in which PC-1 gene can be stably expressed is established and overexpression of PC-1 gene is not sufficient for causing obesity, insulin resistance and type 2 diabetes.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2005年第6期554-556,共3页
Chinese Journal of Endocrinology and Metabolism
基金
国家杰出青年科学基金资助项目(39925023)
国家863计划资助项目(2001AA216081)
上海市科技发展基金资助项目(99JC14029)
