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Construction of Shkbp1 and Other Strains of Genetically Engineered Homozygous Mice

Author: HeXiaoDong
Tutor: LiWeiDong；WangLiJing；GengJianGuo
School: Guangdong College of Pharmacy
Course: Human Anatomy and Embryology
Keywords: genetically engineered mice homozygous Shkbp1 SGTα nef Rip-hSlit2
CLC: Q78
Type: Master's thesis
Year: 2011
Downloads: 26
Quote: 0
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Abstract


Background & ObjectiveCurrently, genetically engineered mice of human diseases model produced in genetic engineering have become an important method and means to study human diseases (including communicable diseases, cancer, cardiovascular diseases, metabolic diseases, age-related diseases, mental diseases and genetic diseases etc.). Compared with normal mice, genetically engineered mice have the un-compared dominance that can simulate the naturally occurring process of human disease in vivo and is more conducive to clinical research and pharmaprojects. It also can carry on studying the mechanism of the disease to the molecular level and is more conducive to the discovery of new drug targets.Crossing genetically engineered mice with spontaneous disease mouse model to study the role of genes in disease, is the most advanced technology used to study the functional genes of human major diseases. In order to study the functions of refered genes in inflammation, cancer and other diseases more accurately, make full use of the great advantages of genetically engineered mouse models in life science research, four strains of genetically engineered mice were constructed by our laboratory, Shkbp1 knockout mice, SGTαknockout mouse, nef1 knockout mouse and overexpression of Slit2 protein Rip-hSlit2 pancreatic mice respectively. Genetic methods were used to obtain a stable genetic background homozygous mice, hopefully to find the roles of other genes related to inflammation, cancer and the role of vascular disease by crossing to ApoE KO, Rip1-Tag2, ApcMin, TRAMP and MMTV-PvMT mice and other diseased mice model.Method1 Breeding genetic engineering miceGenetically engineered mice were housed in SPF-class environment, according to the need for breeding, allocating mice timely after lactation, to make preparation for the screening of homozygous mice.2 The establishment of stable genetically engineered mice and screening of homozygousApplied classic Mendelian laws in genetics to breed the transgenic mice, and combined with PCR to detect the target genes.3 Shkbp1 gene on tumor growthMaking Shkbp1 homozygous knockout mice as the experimental group, wild-type 129 mice as the control group to establish subcutaneous B16 melanoma tumor model Measuring the tumor volume, tumor weight,to further investigate the effect of Shkbp1 knockout mice on tumor growth and metastasis.Results1 129 generation mice cross with F2 positive mice to detect the genotypes of their offspring. when one male and three female Shkbp1 mice have been selected, the offspring gene identification results was 100%; when two male and one female SGTαmice have been selected, the offspring gene identification results was 100%; when one male and one female pVA-hCD2-Nef in mice have been selected, the offspring gene identification result was 100 %. Rip-hSlit2 mouse filter is still in progress.2 Shkbp1 in subcutaneous tumor and metastatic tumor mice model (the results are in statistics)Conclusion1 Shkbp1 homozygous knockout mice were successfully constructed, and it is verified the effect of Shkbp1 gene in tumor growth and preliminary metastasis.2 SGTa homozygous knockout mice were successfully constructed. Generation F2 pVA-hCD2-Nef and generation F1 Rip-hSlit2 TG mice were constructed.

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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