Total RNA was prepared from pituitary gland of new butchered sika cDNA was synthesized by RT\|PCR and used as the template in PCR amplification for β\|subunit gene of sika follicle stimulating hormone(FSH). The PCR p...Total RNA was prepared from pituitary gland of new butchered sika cDNA was synthesized by RT\|PCR and used as the template in PCR amplification for β\|subunit gene of sika follicle stimulating hormone(FSH). The PCR product was 390 bp in 1\^2% agarose gel electrophoresis which just was of the same length as the predicted FSH β\|subunit gene. Cloned to pMD\|18\|T\|Vector, selected 2 positive recombinants at random and analyzed its sequence. The comparison of this amino acid sequence of FSH β\|subunit with some other mammalian showed that it has higher homology with horse, pig, dog and lower homology with rat. Although FSH β\|subunit gene shows species specificity, it is quite conserved among mammalians.展开更多
运用SYBR Green I荧光定量RT-PCR方法和放射免疫方法(RIA)分析了外源性雌二醇(estradiol-17β,E2)对异育银鲫脑垂体-甲状腺轴的影响。实验设计时间为10周,实验组每2周注射一次0.5mg.kg-1体重的E2(用57%酒精溶解),对照组注射同剂量的57%...运用SYBR Green I荧光定量RT-PCR方法和放射免疫方法(RIA)分析了外源性雌二醇(estradiol-17β,E2)对异育银鲫脑垂体-甲状腺轴的影响。实验设计时间为10周,实验组每2周注射一次0.5mg.kg-1体重的E2(用57%酒精溶解),对照组注射同剂量的57%酒精。实验期间没有投喂饲料。在实验条件下,SYBR Green I荧光定量RT-PCR分析结果显示,注射3和5次外源性E2后,促甲状腺激素β亚基mRNA的表达水平与对照组相比均未有显著变化,但随着注射次数的增加,总体呈现升高的趋势。另一方面,RIA结果显示,注射3次外源性E2后,血清中甲状腺激素T3和T4含量水平与对照组相比均显著升高(P<0.05);注射5次后,与对照群相比差异极显著(P<0.01)。以上结果提示,外源性E2影响异育银鲫的正常生理状态。展开更多
文摘Total RNA was prepared from pituitary gland of new butchered sika cDNA was synthesized by RT\|PCR and used as the template in PCR amplification for β\|subunit gene of sika follicle stimulating hormone(FSH). The PCR product was 390 bp in 1\^2% agarose gel electrophoresis which just was of the same length as the predicted FSH β\|subunit gene. Cloned to pMD\|18\|T\|Vector, selected 2 positive recombinants at random and analyzed its sequence. The comparison of this amino acid sequence of FSH β\|subunit with some other mammalian showed that it has higher homology with horse, pig, dog and lower homology with rat. Although FSH β\|subunit gene shows species specificity, it is quite conserved among mammalians.
文摘运用SYBR Green I荧光定量RT-PCR方法和放射免疫方法(RIA)分析了外源性雌二醇(estradiol-17β,E2)对异育银鲫脑垂体-甲状腺轴的影响。实验设计时间为10周,实验组每2周注射一次0.5mg.kg-1体重的E2(用57%酒精溶解),对照组注射同剂量的57%酒精。实验期间没有投喂饲料。在实验条件下,SYBR Green I荧光定量RT-PCR分析结果显示,注射3和5次外源性E2后,促甲状腺激素β亚基mRNA的表达水平与对照组相比均未有显著变化,但随着注射次数的增加,总体呈现升高的趋势。另一方面,RIA结果显示,注射3次外源性E2后,血清中甲状腺激素T3和T4含量水平与对照组相比均显著升高(P<0.05);注射5次后,与对照群相比差异极显著(P<0.01)。以上结果提示,外源性E2影响异育银鲫的正常生理状态。
