转pGH基因猪外源基因染色体定位研究

应用地高辛标记探针对转pGH基因(猪生长激素基因)猪染色体进行原位杂交,经胶体金抗体、银增强放大系统检测外源pGH基因在染色体上的整合位点。研究表明,转基因阳性个体间外源基因整合位点存在差异,但对个体而言,外源基因总是集中分布于某一特定的染色体上。本研究将为探究外源基因整合位点与其表达效率的关系及今后定点整合的研究提供理论指导。

Exogenous pGH Gene Localization on Chromosomes of the Transgenic Pigs

The pUC19-OMT plasmids were cut by Ssp 1 and the products in 900 bp were recovered from low-melting agarose gel and used as probes which were labeled by Digoxigenin. After being denatured, the probes were dropped on the chromosome samples which were also denatured to anneal with them. The anti-digoxigenin antibodies labeled with colloidal gold were used to act with the chromosome samples. In order to localize the exogenous pGH genes(porcine growth hormone gene) on chromosomes detected with optical microscope and improve the sensitivity, digoxigen gold signals are amplified by silver precipitation. After calculating the number of silver grains on every chromosome under the optical microscope, we analyzed the data with statistical methods. The results show that the integrating sites of exogenous pGH genes are very different among the positives. However, it is clear that the exogenous genes in one are always of the tendency to integrate in one specific site on a certain chromosome. These data are of great significance for studying the site-specific integration and the expression efficiency of exogenous genes in the future research.