| The transgenic cloning technology is used, it can change the genes of an organism in ways not possible through traditional breeding techniques providing opportunities for new plant varieties and animal breeds, to the breeding of transgenic cloning technology is a long process to found a new choiceness traits plant and animal breeds. At present, the researchers has been successfully used transgenic cloning technology to produce all kinds of animals, such as sheep, cows, pigs, rabbit, rhesus monkey etc. Cloning technology is product a population for scientific research in a short time, shorten the breeding time. Transgenic cloning technology has been widely used in various fields, that is a powerful tool to offers gigantic benefits for new varieties breeding, treatment of human disease.1. Induction of experimental type I diabetes in cloned Wuzhishan miniature pigs by alloxan. In order to explore the cloning animal models of diabetes in the diabetes treatment and application research. Induction of experimental type I diabetes in cloned Wuzhishan miniature pigs by different concentration of the alloxan. In this study, T I D (type I diabetes) was experimentally induced in cloned Wuzhishan miniature pigs with alloxan. In brief, an intravenous injection of alloxan (group B:170mg/kg, n=3) was administered to pigs weighing between27and39kg. Sterile saline was administered to control pigs (n=3). We determined the glycometabolism related index, performed an intravenous glucose tolerance test, and carried out immunohistochemistry experiments. There were no significant differences in body weight, blood glucose, and serum insulin in all groups, before treatment. The level of blood glucose was significantly higher (P<0.05) in group B (12.18±0.70mmol/L) than in the control (2.93±0.39mmol/L). By contrast, the level of serum insulin was lower in group B (5.641±0.573(?)IU/mL) than in the control (7.578±0.539(?)IU/mL). Histological studies by hematoxylin and eosin (H&E) revealed a loss of β-cells in the pancreas from pigs treated with170mg/kg alloxan. Immunolocalization studies showed a decrease in insulin reactivity in this treatment group as well. To conclude, our model holds promise in future studies of diabetes drug testing and islet xenotransplantation.2. Genetic performance and transgene expression of Red Fluorescent Protein (RFP) transgenic Wuzhishan miniature pigs and offsprings. Analysis of the blood physiological indexes and mRFP expression of transgenic Wuzhishan miniature pigs of the hybrid progeny. This study to explore the reproductive capacity of red fluorescent protein transgenic Wuzhishan miniature pig, and its stability of exogenous gene expression. Methonds of this rearch:Generation of transgenic Wuzhishan miniature pigs, fibroblasts were collected from the ears of adult Wuzhishan miniature pigs to build a donor cell lines, exogenous gene expression of the pCX-mRFP will was transfected into fibroblasts. Then wuzhishan miniature pigs was cloned thought somatic cell nuclear transfer (S C N T) technology, transgenic wuzhishan miniature pigs nowborn with generated fluorescent protein marker, mRFP (monomeric red fluorescent protein)—F0. The transgenic pigs expressed mRFP in all organs and tissues at high levels. MRFP1expression of transgenic Wuzhishan miniature pig hybrid intercourse with wild type non transgenic Wuzhishan miniature pig. Generation of the hybrid progeny—F1with generated fluorescent protein marker, mRFP and analysis of the mRFP and the blood routine. Result:the positive mRFP Wuzhishan miniature pigs F1were compared with the FO have a consistent in characteristics and appearance,F1generation’s appearance show up a bright pink,.Analysis of blood samples showed no significant difference,F0is compared with F1blood physiological indexes.These results demonstrate that F1generation mRFP genetic stability and effectively expressed in the tissues. F1generation of Wuzhishan miniature pigs efficiently express mRFP can be generated. The transgenic animals as excellent models may have applications in medicine and agriculture.We produce monomeric red fluorescent protein mRFP1transgenic pigs, the red fluorescent protein gene, can stable genetic to F1offspring,and highly expressed. In the future we use cross breeding technology to expand the number of red fluorescent protein transgenic wuzhishan miniature pig, and set up a new population. The now variety will be applied to study the treatment of cancer, the purpose gene tracking, production of animal disease model, etc,and due to its small size, bright appearance, beautiful and easy to feed, also can be used as a new fashion pets. There are broad development prospects of scientific research and market economy. |
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