Main works and their important results are showed as following . firstly , the cdna encoding bullfrog gh is amplified by use of the total rna , extracted from adult bullfrog ' s pituitary , as the template , pi ( 5 ' - cggatcc atg gct tca ggg tta ggc ) and p2 ( 5 ' - cgaattc tta aaa ggt gca gtt gct ) as the primer pair and one particular cdna product , 660bp , was obtained after the rt - pcr - the product was purified by using dna agarose gel extraction method . after the purified cdna and pmd18 - t vector were ligated at 16 over night , the ligation products were transformed into e . coli strain dh5a and then the transformants were screened by clone pcr method 首先，以成体牛蛙的脑垂体总rna为模板，进行rt ? pcr扩增得到约660bp的特异性pcr产物带，切下琼脂糖凝胶中的特异产物带进行cdna胶回收，将cdna与pmd18 ? t载体进行t ? a克隆连接并转化到dh5 e . coli后，进行菌落pcr 、质粒酶切鉴定，筛选出阳性菌株，测序结果经blast分析，与已报道的牛蛙生长激素基因高度同源，证实此阳性克隆为牛蛙生长激素基因转化子，命名为pbfgh 。
In this study , the stem segments of new shoot with axillary buds of well - growth tetraploid black locust trees were used as explants . the effects of different basic mediums , different hormone kinds and their concentrations ratios , different sucrose concentrations on calli induction , buds differentiation and rooting in the process of establishment of high frequency regeneration system of tetraploid black locust were studied . on the base of high frequency regeneration system , the effects of various factors on transformation efficiency of badh mediated by agrobacterium tumefaciens were discussed in the light of gus histochemical assays 本实验首先以生长良好的四倍体刺槐优株上当年生新梢的带腋芽茎段为外植体，研究了在四倍体刺槐高频再生体系的建立过程中不同基本培养基、不同激素浓度及其配比、不同蔗糖浓度对愈伤组织的诱导、芽的分化及生根的影响；然后在得到高频再生体系的基础上，通过农杆菌介导法转化甜菜碱醛脱氢酶( badh )基因，以gus染色组织分析为依据探讨了影响转化效率的各种因素，建立了高效、可重复的基因转化体系，为四倍体刺槐目的基因的导入打下了基础。
The recombination gene micyoinjected is 5nl dna ( about 106 - 107 ) into everyone oocytes or cytoplasm of grass carp . there were 32197 oocytes of 12 series via micyoinjection and achived 12945 fries in 2000 year , by hatching and raising . genomic dna was extracted from grass carp who were transformated in 1120 grass carp presented postive reaction by blood 实验通过将hu - - ifn基因重组分子采用显微注射法将这种重组基因转化草鱼1 - 2细胞期的受精卵，即以narashige显微注射仪，将5nldna (约10 ~ 6 10 ~ 7分子)直接注射到选择好的受精卵核的动物极上。
