By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A...By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A(♀♂) and 2B(♀♂)transgenic homozygote mice, and 6 bi-heterozygote mice(4♀2♂) by mating A and B homozygote mice were taken. Blood serum of these transgenic mice and their off-springs’ were collected, and HBV DNA expressing in blood serum of transgenic mice was tested by polymerase chain reaction and ELISA (PCR- ELISA). The results were as follows: HBV DNA expression in the blood of bi-heterozygote mice was much more than that of homozygote and heterozygote mice. Breeding bi-heterzygote mice is likely to be a good way of high expression of foreign gene in transgenic animals.展开更多
[Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DN...[Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DNA in non-GMO soybeans as the endogenous reference standard,the method of gradient dilution was used for separately calculate Ct value of endogenous reference gene and plasmid DNA and correlation standard curve equation of logarithm of copies,and then to calculate the copies of samples through substituting thus-obtained Ct into the standard curve equation.[Result] The standard curve equation of endogenous reference gene is y=-3.422x＋35.201,R2=0.998;and the standard curve equation of exogenous gene is y=-3.348x＋34.890,R2=0.999.Nos terminator and its lower boundary sequences in transgenic soybean is of single copy.[Conclusion] The study has provided a theoretical basis for determining exogenous gene copies in transgenic soybean.展开更多
文摘By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A(♀♂) and 2B(♀♂)transgenic homozygote mice, and 6 bi-heterozygote mice(4♀2♂) by mating A and B homozygote mice were taken. Blood serum of these transgenic mice and their off-springs’ were collected, and HBV DNA expressing in blood serum of transgenic mice was tested by polymerase chain reaction and ELISA (PCR- ELISA). The results were as follows: HBV DNA expression in the blood of bi-heterozygote mice was much more than that of homozygote and heterozygote mice. Breeding bi-heterzygote mice is likely to be a good way of high expression of foreign gene in transgenic animals.
基金Funded by Program of Technology Bureau of Harbin(2010RFQXN101)Sub-project of Transgenic Significant Specific Project(2008ZX08012-001)~~
文摘[Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DNA in non-GMO soybeans as the endogenous reference standard,the method of gradient dilution was used for separately calculate Ct value of endogenous reference gene and plasmid DNA and correlation standard curve equation of logarithm of copies,and then to calculate the copies of samples through substituting thus-obtained Ct into the standard curve equation.[Result] The standard curve equation of endogenous reference gene is y=-3.422x＋35.201,R2=0.998;and the standard curve equation of exogenous gene is y=-3.348x＋34.890,R2=0.999.Nos terminator and its lower boundary sequences in transgenic soybean is of single copy.[Conclusion] The study has provided a theoretical basis for determining exogenous gene copies in transgenic soybean.
