1.Construction And Expression Of TIMP-1 Transgenic Mice Model 2.Chromosomal Location Of TIMP-1 Transgenic Mice With FISH

Matrix metaloproteinases (MMPs) and it's inhibitor tissue inhibitors of metalproteinases (TIMP-1) is a very important system in balance between the synthesis and degradation of extracellular matrix (ECM). While TIMP-1 is a major inhibitor in degradation of ECM and has been generally accepted as a key factor in ECM remodeling and in the pathogenesis of progressive renal diseases. Interestingly, besides its MMP-inhibiting activity, TIMP-1 also has some other effects, such as erythroid potentiating activity and cell growth promoting activity in a wide range of cells, suppressing apoptosis in B cells and breast epithelial cells. This study was designed to examine the role of TIMP-1 in ECM degradation and whether TIMP-1 have effects on the apoptosis and differentiation of renal cells except its function of MMPs' inhibition.To further investigate the effects of overexpression of TIMP-1 on kidney in vivo, we constructed transgenic mice for TIMP-1 by microinjection. The TIMP-1 transgenic mice were confirmed by PCR assay and Southern-blot analysis . In experiment 1200 ovum were injected into pronuclear at 1-cell egg stage, 896 of which were alive . living rate is 74. 7%. 612 eggs injected were then implanted into the oviduct of 21 pseudopregnant females.10 of 21 pseudopregnant females were pregnancy. 32 pups were produced.28 of which were alive. The birthrate was 5.2%. Fourfounders were identified(4/32;12. 5%).Male founder was mated with normal female mice and produced 100 offsprings as the 1st generation. By PCR and Southern blot test positive mouse were 17 and 7 respectively. By this method the TIMP-1 transgenic mice have been bred successfully and have progressed to 5th generation. All of the 5th generation was positive mouse,, The positive reactions were mainly observed in the organs such as liver-. spleen^ kidney^ lung and brain by Northern blot.