t vector造句

• Then the pcr product was purified , ligated into pgem - t vector by ta cloning
  筛选阳性克隆，测序，大量制备序列完全正确的质粒。
• The pcr product was cloned into pmd18 - t vector . the positive recombinant clone was identified by pcr and endonuclease digest
  提取重组质粒经pcr鉴定和酶切鉴定后，对插入片段进行序列测定及分析。
• The product of pcr named vp6 is approximate 1 . 3kb in length . the vp6 gene was cloned into pmd18 - t vector and sequenced
  将其插入克隆载体质粒pmd18 - t的ecorv酶切位点处，构建重组质粒pmd18 - t ? vp6 。
• Pgem - t vector system , jm109 competent cells , reverse transcription kit and in vitro translation kit were purchased from promega company
  Pgem一t载体，感受态细菌jm109 ，反转录试剂盒和体外翻译试剂盒均购自promega公司。
• 2 . pcr products can be purified by ctab method , which removing the dntp and primers . the purified products can be used in the ligation reaction to pmd18 - t vector with high efficiency
  2 、采用ctab进行pcr产物的纯化，以去除pcr产物中的引物和dntp ，所得产物可用于与t载体连接反应，以提高连接效率。
• 2 . using a pair of degenerate primers based on the conserved region , hmg - box , of human sry gene , eight different fragments were amplified from both female and male rana rugulosa wiegmann then cloned by using pmd18 - t vector and sequenced
  2 、参照人sox基因设计了一对兼并引物，扩增了虎纹蛙的sox基因，并对扩增产物进行了克隆和测序。
• To device a primer pairs and amplify the full length pstvd by rt - pcr , the positive rna extraction from tuber sample was used as template . the product of rt - pcr was purified and connected to the plasmid pmd 18 - t vector
  Cdna核酸斑点杂交反应( nash )检测pstvd方法准确、灵敏度高，一次检测样品数量多，且对于异地样品检测非常方便，是以往其它检测方法的有效补充。
• Two degenerate primers were designed and synthesized according to the highly conservative sequences among the known - glc genes . a cdna fragment of 208bp was amplified by rt - pcr , which was subsequently cloned into pmd18 - t vector for sequencing analysis
  利用genbank中已经登录的其它植物中该酶基因的保守序列，设计一对简并引物，采用rt - pcr技术，从茶树扩增出208bp的cdna片段。
• The tmek2 and tmek2mut was amplified by pcr and then cloned to pmd 18 - t vector . to express the genes in plants , the coding region of tmek2 and tmek2mut were placed between the 35s promoter and nos terminator of pibl , respectively
  载体构建的过程包括目的基因的pcr扩增、目的基因连接到pmd18 - t载体、目的基因连接到中间表达载体pib1 、目的基因及启动子、终止子连接到表达载体pbin19 。
• Designing a specific primer pair based on the sequence of the gene ubia in e . coli mc4100 , a dna fragment was amplified from the genomic dna of e . coli mc4100 by pcr and then was cloned into pucm - t vector . the cloned fragment was sequenced
  根据e . colimc4100的ubia基因全碱基顺序，合理设计引物，以e . colimc4100基因组dna为模板进行pcr扩增，将pcr产物克隆于pucm - tvector后对其进行测序鉴定。
• Clone human wnk4 full length cdna into pgem - t vector human kidney total una was extracted and used as template to amplify wnk4 full length cdna with the forward primer ( 7 - 27 ) and reverse primer ( 3808 - 3833 ) with the long template expanded pcr system kit
  5 .原位杂交以小鼠肾脏总翩a为模板，扩增wnki基因外显子1 ， 24一28 ，以a片段和wnk4基因外显子13一16片段，纯化后连接在pgem一t载体上。
• 2 . cloning of the pcr products the pcr products were purified by agarose gel electrophoresis and was ligated with pucm - t vector . by the method of pcr and enzyme digest analysis . the result shows that the plasmid containing cpti gene was transferred into e . coli dhso
  Pcr产物的克隆采用a / t克隆法，将pcr产物经琼脂糖凝胶电泳纯化回收后用t4连接酶与pucm - t载体连接，构建成克隆载体puc - cp ，转化大肠杆菌dh _ 5 。
• Pcr methodology was adopted for cloning of chitinase encoding genes . based on the sequences of chitinase gene from genebank , the primers for chitinase gene amplification were designed . pcr fragment was ligated with pmd18 ~ t vector and transformed into e . coli dh5 a
  尽管同源性比较低，但酶活性检测发现该基因片段具有几丁质酶活性，认为此pcr片段含有几丁质酶编码基因的全序列或部分序列，此基因片段是一个新基因或基因片段。
• The library was rescued with phage m13k07 in order to display scfv on the surface of the phage and to form the recombinant phage antibody library . one of positive scfv clones , named pcsal , was selected with phage - elisa after panning and screening by bull sperm three times . scfv fragment , amplified from pcsa1 , was ligated to pmd18 - t vector for sequencing analysis
  取阳性重组噬菌体抗体克隆株pcsa1 ， pcr扩增其scfv基因，筛选重组子进行序列测定，发现其序列符合小鼠抗体基因的一般特征，并且与几株抗磷酸胆碱的抗体重链和轻链可变区序列的同源性达80以上；推测pcsa1scfv针对的抗原是磷酸胆碱类物质。
• After electrophorised on 1 % agarose gel , the pcr production was purified with agarose gel dna extraction kit . the segment was ligated with vector pmd18 - t and then was tranformed into the competent cell of dh5 a . a construction mstnd - pmd18t was generated by inserting the sequence of 254bp into pmd18 - t vector and selecting the sense clones . positive clone was identified by three ways : endonuclease digestion , pcr and sequencing . the result showed that the cloned sequence coincides with the designed sequence . this construction was digested with nco i and xho i and ligated the pet28a ( + ) vector digested with the same enzymes using dna ligation kit . the production of ligation reaction was transformed into the competent cell of bl21 ( de3 ) . after 12 - 16 hours of culture , several colnes appeared on the plate . some positive clones were selected to extract their plasmid . these plasmids were digested by nco i and xho i and indentified by pcr . a contraction , mstnd - pet28a was generated . the result showed that the cloned sequence coincides with the designed sequence
  F _ 1长38bp ， r _ 1长36bp ，其它片段均40bp长， f _ 1和r _ 1片段两端分别加上限制性内切酶nco和xho的识别位点序列。用成对单链片段进行延伸反应，然后用其他单链片段作为引物，进行pcr扩增，用dna快速纯化回收试剂盒回收所得254bppcr产物，与pmd18 - t载体连接、转化dh _ 5 。受体菌感受态细胞，利用蓝白斑遗传学筛选法筛选阳性克隆，提取其质粒，采用nco和xho双酶切鉴定，获得了254bp的片段；用pmd18 - t载体上的特异引物rv - m和m13 - 47进行pcr鉴定，获得300bp的片段。
• Part two : studies ; l . the sox gene of dinodon refozonatum was amplified by using a pair of primers which can amplify the conservative motif ( hmg - box ) of human sry gene . the amplification band was observed in both male and female dinodon refozonatum , whose length was consistent with that of human sry gene , which about 220bp . the result of sscp analysis showed that there were many differences in the sox gene sequence between dinodon refozonatum and human , and there was a few differences between male and female dinodon refozonatum . 2 . using a pair of degenerate primers based on the conservative region , hmg - box , of human sry gene , six different fragments were amplified from either female or male dinodon refozonatum , then cloned by using pmd18 - t vector and sequenced
  结果显示： ( 1 )赤链蛇基因的扩增片段与人sry基因扩增片断大小相同，为220bp左右； ( 2 )雌、雄赤链蛇sox基因的扩增片段大小虽然与人的相同，但其单链迁移率与人的有较大差异，而且雌雄个体间有明显差异，预示该基因的dna序列在雌雄个体中可能有差异； 2 、参照人sry基因hmg - box保守区的序列，又设计一对兼并引物，扩增了赤链蛇的sox基因，并对扩增产物进行了克隆和测序。
• The pcr products were purified and cloned into pucm - t vector then vector pp - rd29a was constructed . by white - blue screening , colony pcr and enzyme digest analysis , the result showed the plasmid containing p - rd29a fragment was transferred into e . coli xl 1 - blue . the sequencing of the single recombinant bacterium containing the inserted p - rd29a fragment was carried out by baiasia bio - engineering co . ltd
  纯化的目标产物经taqdna聚合酶加a碱基后，采用taclonging方法克隆到pucm - t载体中，得到重组质粒pp - rd29a ，转化大肠杆菌xl _ 1 - blue ，挑选白色的重组子菌落进行colonypcr筛选和质粒的酶切验证。
• The chi gene fragment was amplified by means of rt - pcr , using flower mrna of saussurea medusa maxim , as template , and a pair of specific oligonucleotides ( according to the chi gene sequence , af509335 ) as primer , and cloned into pucm - t vector with ta connection . the vector was named pucm - chi
  Chi基因的正向和反向植物表达载体的构建用ecor和sal双酶切p ~ ( ucm - chi )和中间载体ptriplex2 ，回收、连接目的片段与载体，构建中间载体p ~ ( tripchi ) ，选择正向重组质粒。
• A complete cdna of annfaf expressed specifically in strawberry fruit was cloned via rt - pcr and was subcloned to pmd18 - t vector to construct the recombined pmd18 - t ( annfaf ) vector . the inserted fragment was sequenced and the character of deduced amino acids was analyzed to obtain the antiserum
  )为试材，从其完熟果实中提取总rna ，以genbank中发表的annfafcdna序列为依据，利用rt - pcr技术克隆得到草莓果实特异表达基因annfaf的cdna序列，将其克隆到pmd18 - t载体中构建成重组克隆质粒pmd18 - t ( annfaf ) 。
• The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector . the sequencing results showed that pap gene had 99 . 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ) . the iptg - inducible expression vector containing the pap gene was constructed and transferred into e . coli bl21 ( de3 ) - plyss
  将缺失型pap基因克隆到植物表达载体pbi121中，通过液氮冷冻法将重组质粒转入农杆菌lba4404细胞中，然后采用叶盘法，在该农杆菌的介导下将pap基因导入普通烟草中，经过卡那霉素抗性筛选，最后获得了转pap基因的工程烟草植株，摩擦接种烟草花叶病毒( tmv ) ，与非转基因烟草相比，能够推迟症状表现达2月之久，说明pap基因能够在其它植物体内产生有活性的高抗病毒的蛋白质。