The fermentation liquid of the transformant 轉(zhuǎn)化子發(fā)酵液也具有脫毛效果。
In contrast , most of the pollen grains in the non - transformant have plump shape and were stained into red color uniformly 轉(zhuǎn)正向表達(dá)載體在煙草正常花粉粒的百分比較對(duì)照植株降低了42 。
The transformant would flower in 8 to 10 weeks , sometimes under in vitro culture , after the leave dies was infected by agrobacterium tumefaciens 轉(zhuǎn)化植株從浸染起8 - 10周可以開(kāi)花,甚至在培養(yǎng)瓶中即可完成其生活史。
And another change was that the spore of the transformant was difficult to release , p3a was not useful in genetically engineered strain construction in this way 此外, pcr擴(kuò)增發(fā)現(xiàn)轉(zhuǎn)化子比出發(fā)菌ybt - 1520多了cry1ab的特異帶, sds - page電泳證實(shí)cry1ab的存在。
Confirmation of transformant . integration of sag - ipt into chrysanthemum genome was confirmed in two thirds of the putative tansgenic plantlet be pcr amplification 通過(guò)pcr擴(kuò)增表明,轉(zhuǎn)基因植株中可以檢測(cè)出有目的基因( ipt基因)的存在,目的基因已成功地插入菊花的基因組中。
Transformation was done by electroporation . human fl extracellular domain cdna transformed to yeast host strain km71 , then his + muts phenotype transformant was screened out and cultured in flasks , and rhfl was expressed under the induction of 0 . 5 % methanol 我們提取了km71ppic9k - fl轉(zhuǎn)化菌株的基因組dna進(jìn)行southern實(shí)驗(yàn),檢測(cè)目的基因的整和插入;提取總rna ,進(jìn)行了northern實(shí)驗(yàn),檢測(cè)fl基因在轉(zhuǎn)化菌株中的表達(dá)。
Transformant ybt - 833 - 1 kept all indigenous plasmids , ybt - 833 - 2 lost a plasmid carrying crylab and ybt - 833 - 3 lost all indigenous plasmids carrying icp genes . comparing their potency with natural strain ybt - 833 , the overall toxicity of all transformants was not noticeably higher than strain ybt - 833 質(zhì)粒pbmblc電脈沖轉(zhuǎn)入菌株ybt - 833 ,得到了幾個(gè)icp基因背景不同的轉(zhuǎn)化子,它們的綜合毒力較出發(fā)菌ybt - 833都沒(méi)能得到提高。
The hwtx - i gene was chemically synthesized according to its known cdna sequence , the gene was inserted into vector ppic9k which contained aoxj promotor and the sequence of a secreting signal peptide - a - factor , the cloning ppic9k / hwtx - i was constructed and confirmed by two - step pcr and dna sequence analysis , then it was transformed into host strain gs115 , a his + muts cell line was screened and multicopy transformants were screened by various g418 concentrations , the multicopy transformant was named gh1 . gh1 was cultivated in flasks . after 6 days of induction by 0 . 5 % methanol , the supernatant was checked by 16 . 5 % tricine - sds page , which showed there was a band in the position of 3 . 5 - 6 . 1kd , then it was isolated and desalted by ultrofiltration followed by ion exchange of cm column , after reverse phase hplc of ci8 and vacuum drying , the purified rhwtx - 1 was obtained which was proved to be correct recombinant hwtx - i by tricine sds - page , maldi - tof mass spectrometry , amino acid composition analysis , the n - terminal amino acid sequence and its biological activity , the final field of the purified rhwtx - i was about 80mg / l , accounting for 23 . 6 % of it total secretory proteins 將帶有hwtx -基因的ppic9k經(jīng)blgii線性化后,轉(zhuǎn)化酵母宿主菌gs115原生質(zhì)體后經(jīng)篩選陽(yáng)性克隆并經(jīng)表型鑒定為his ~ + mut ~ s酵母菌,進(jìn)一步用遺傳毒素g418篩選多拷貝的轉(zhuǎn)化菌株,命名為gh1 ;將gh1甲醇酵母菌用0 . 5的甲醇誘導(dǎo)表達(dá),發(fā)酵上清經(jīng)90飽和度的( nh _ 4 ) _ 2so _ 4沉淀, yw - 3 ( mwc03000 )的超濾膜超濾,再經(jīng)cm陽(yáng)離子交換, c _ ( 18 )反相hplc純化得到分子量為4kd左右的組分,其中4289 . 05的組分經(jīng)質(zhì)譜鑒定,氨基酸組成分析和序列測(cè)定為正確的表達(dá)產(chǎn)物,生物學(xué)活性表明其活性為天然毒素活性70 % ,表達(dá)量為80mg / l 。
Plant expression vector lba4404 ( pbin + - 35s - cbf3 + pal4404 ) was used in the genetic transformation of cucumber and micro - tomato . establishment of high efficiency regeneration system and determination of parameters involved in agrobacterium - mediated transformation , screening and characterization of transformant were systemically researched 將植物表達(dá)載體pbin ~ + - 35s - cbf3對(duì)黃瓜和小番茄進(jìn)行遺傳轉(zhuǎn)化,建立了黃瓜高頻再生和遺傳轉(zhuǎn)化體系,獲得了一株轉(zhuǎn)基因黃瓜植株,并通過(guò)了植物基因組pcr檢測(cè);獲得了兩個(gè)具有kan抗性的小番茄再生綠芽。
T . identification of charactrization of transgenic mustard plants the putative transformant regeneration plants were assayed by pcr and pcr - southern blot analysis . both analysis the target bands were observed . so the integration of the cpti gene into mustard genome dna was confirmed . the result of insect - resistance showed that the transgenic plants are more resistant than non - transgenic plants 轉(zhuǎn)基因芥菜植株的鑒定取再生苗的葉片提取dna ,進(jìn)行pcr擴(kuò)增和pcrsouthernblot分析,轉(zhuǎn)化再生植株大部分呈陽(yáng)性,而非轉(zhuǎn)化的再生植株均為陰性,證明cpt基因已存在于芥菜基因組中。