6 . transformation system of mustard a serials of kanamycin concentration was added to optimum medium to test the explants resistance capacity of two kinds of mustard . the transformation procedures described were derived from numerous regeneration and trasformation designed to test factors that might affect shoot regeneration , which including length of co - cultivation . those producing the best result parameters were described as below : after the mustard explants were precultured on regeneration medium for 2 days . they were inoculated with agrobacterium for 20 minutes . inoculated explants were co - cultivated for 4 days and in shadow at first 2 days . then transferred to the same medium plus 30 mg / l kanamycin and 500mg / l garb . all of them were transferred to fresh medium every 2 weeks . the kan - resistant plants were regenerated 芥菜外植体高频遗传转化体系的建立在最适培养基上试验了两类芥菜的三种外植体对卡那霉素的敏感性、预培养天数、浸菌时间等因素的影响，建立了芥菜高频转基因再生体系：取生长4天的芥菜子叶、下胚轴和25天的叶片在分化培养基上( ms + ba3 . 0mg / l + naa0 . 1mg / l )预培养2 - 3天后，投入农杆菌菌液中浸染20分钟，在分化培养基上暗培养2天，正常条件下培养2天后，转入抗性培养基（ ms ba3
Transformed shoots were selected on solidified medium containing kanamycin . ten kanamycin resistant transformants were obtained by direct or induction calla . these transformants were checked by pcr , pcr - southern blot and southern blot , confirmed that two positive transformants were integrated into the genome of boechmeria nivea l guad 对所得到的抗性转化株进行了pcr 、 pcr - southernblot 、 southernblot检测，其中2株呈阳性，证明vp4基因已整合到苎麻基因组dna中。
The chloroplast shsp gene was screened from the cdna library of tomato flower by pcr strategy and confirmed by sequencing . but difference was found at 3 bases of the sequence from the reported in genbank . then , an integrated vector prok ii of the chloroplast shsp gene and nptii gene ( a kanamycin resistant gene ) with camv35s promoter was constructed and introduced into tomato mediated by agrobacterium tumefaciens lba4404 . transgenic tomato were screened by their ability of growing on media containing kanamycin 本实验采用pcr方法从番茄花cdna文库中克隆到叶绿体shsp基因，经测序证实与genbank中已发表的序列在编码区相差2个碱基，其中一个碱基导致1个氨基酸的改变。将叶绿体shsp基因定向克隆于带有组成性表达启动子camv35s的植物表达载体prok中，冻融法转化农杆菌lba4404 ，利用叶圆盘法对番茄进行ti质粒介导的遗传转化。
Establishment of carrot genetic transformation an efficient transformation protocol based on agrobacterium tumefacien lba4404 was created in the study . good results were gained when use fresh hypocotyls as infectious explants , co - cultivate in medium supplemented with low concentration of acetosyringone ( 25 m ) , screen in medium with loomg / l kanamycin . it is time saving when anti - culli are fisrt screened in low concentration of antibiotics then transfered to high concentration ones , and remove antibiotics when regenerate 建立了高效的遗传转化体系以pbi121和ptok233为转化质粒，在农杆菌lba4404介导下，对胡萝卜遗传转化体系进行系统研究，首次为该胡萝卜品种建立一套高效的遗传转化体系，结果为:最适转化受体为新鲜下胚轴及经预培养的下胚轴;共培养时，低浓度的乙酞丁香酮( 25pm )对转化具有促进作用;适宜的卡那霉素筛选浓度是loom岁l 。
The resulting plasmid , named prok - sod2 , was mobilized to agrobacterium tumefaciens strain gv3101 used for plant transformation . the yeast sod2 gene was introduced into arabidopsis thaliana ( ecotype landsberg erecta ) by agrobaterium tumefaciens - mediated transformation with floral - dipping method under the control of camv 35s promoter . transformants were selected for their ability to grow on medium containing kanamycin ( 30mg / l ) , several homozygous lines that were all tolerant to kanamycin were selected and used for further molecular and physiological determination 本实验将sod2基因构建到植物表达载体prok中，导入农杆菌后，进行植物遗传转化，实现其在拟南芥中过量表达，在含30mg l的卡那霉素的培养基上筛选获得纯合转基因株系，自交一代获得足够的纯和转基因种子后，对其进行了分子生物学的验证及生理指标的检验。
The results indicated that all isolates exhibited a susceptibility to amikacin and ceftriaxon , and 67 isolates showed a greater or lesser degree resistance to streptomycin , kanamycin , gentamicin , ampicillin , tetracycline , chloramphenicol , florfenical , cefotaxime , cephalothin and ceftiofur , to which 22 isolates exhibited a susceptibility . some isolates showed resistance to multiple antibiotics and displayed a highest level resistance to streptomycin with a frequency of 43 . 8 % , followed by tetracycline with a frequency of 30 . 3 % 结果表明，所有分离菌株均对阿米卡星和头孢曲松敏感； 67株对链霉素、卡那霉素、庆大霉素、氨苄西林、四环素、氯霉素、氟苯尼考、头孢噻肟、头孢噻吩、头孢噻呋表现出不同程度的耐药性，其中对链霉素的耐药率最高，为43 . 8 ，其次为四环素( 30 . 3 ) ，其余22株为敏感菌株。
Our experiment indicates : ( 1 ) the optimal concentration of kanamycin for screening torenia fournier regenerated buds was 400 mg / l . the ideal transformation was obtained in the following conditions : the leaf discs were dipped in agrobacterium suspension that od600 was 0 . 1 for 10 ~ 20 min ; subsequently cocultivated on the ms solid coculture medium containing 20umol / l acetosyringon for 7 to 8 d at 23 , and the induction ratio of regenerated buds was 27 . 2 % 研究结果表明： ( 1 )筛选蓝猪耳转化芽的最适卡那霉素浓度为400mg l 。 od _ ( 600 )为0 . 1的菌液浓度菌液浸染叶盘10 20min ，固体共培养基(含20 mol l乙酰丁香酮)上23共培养7 8d可获得理想的转化效率，转化芽诱导率为27 . 2 ； 16h光照， 8h黑暗是较理想的共培养光周期；茎段是较好的转化受体。
The chimeric gene was introduced into arabidopsis through the method of vacuum infiltration which is widely used in arobidopsis tansformation . transgenic plants were screened by means of kanamycin resistance and further identified by genomic pcr and western blotting , through the phenotype observation of transgenic plants , the speculation that apoplast calmodulin may play a certain role in regulating the development and growth of plants was made 最后，融合基因被插入二元载体plbj21 ，通过电击法转入农杆菌gv3101 ，真空渗入法转入了拟南芥，转基因拟南芥通过kan抗性筛选而得到，并进一步进行了基因组pcr鉴定和western杂交的鉴定。
