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사람 성장호르몬이 유선조직에서 특이적으로 발현되는 형질전환 생쥐의 개발
유대열(D . Y . Yu),이철상(C . S . Lee),강만종(M . J . Kang),한용만(Y . M . Han),이경광(K . K . Lee) 한국축산학회 1993 한국축산학회지 Vol.35 No.1
In an attempt to develop transgenic animals producing biologically active proteins into the milk., expression vectors (pC and pCS) were constructed. The pC vector consists of the promoter 2.8-kb DNA fragment of rat ?-casein gene and pSP70 plasmid. The pCS vector contains the SV40 poly(A) sequences in the multiple cloning sites of pC vector. Human growth hormone gene was then inserted into the cloning site of each vector, which was designated to pChGH and pCShGH. The plasmids were injected into one-cell pronuclear mouse embryos as a purified Xhol/Hpal fragments containing no procaryotic sequences. The injected embryos were implanted into pseudopregnant females, and 117 mice were born. The live of them were identified as being transgenic (ChGH line: 2 mice, CShGH line: 3 mice) by diagnostic Southern blot hybrization with human growth hormone gene. Among them, three transgenic mice transmitted the DNAs integrated on their chromosomes to their progenies and secreted human growth hormone in milk at the level of 160 ng/㎖ (ChGH-2-2 line), 80 ng/㎖ (CShGH-1 line), and 2-4 ng/㎖ (CShGH-3 line), respectively. The above cases show that no human growth hormone was detected in the serum, and these results suggest that the inserted gene is specifically expressed in the mammary gland of the transgenic mice by the regulation of the β-casein promoter.
Transgenic 생쥐생산에 있어서 미세주입시기 및 외래유전자의 농도가 삽입빈도에 미치는 영향
한용만(Y . M . Han),이철상(C . S . Lee),강만종(M . J . Kang),유대열(D . Y . Yu),이경광(K . K . Lee) 한국축산학회 1990 한국축산학회지 Vol.32 No.6
The present study was undertaken in an attepmt to improve the efficiency of integration frequency in transgenic animals. In order to examine the effect of microinjection time on integration frequency, foreign DNA was microinjected into the male pronucleus of each fertilized egg at 2hr intervals from 21 to 27hr after HCG injection. The results revealed that the overall integration efficiency was somewhat higher in the group of 23-25hr than in the other groups. Wealso investigated on the effect of DNA concentrations(2, 4, 8 & 20ng/ul) on integration frequency, but differences between experimental groups were not statistically significant.