The plasmid dna was cut and dephosphorylated . the linearized plasmid dna was ligated to the pseudomonas sp . dna fragments and resulting recombinant plasmids were used to transform escherichia coli xll - blue 提取细菌总dna ，用限制性内切酶部分酶切后，回收2一6kbdna片段，与克隆载体puc18连接，转化大肠杆菌，构建了该菌株的基因组文库。
In this experiment , ri gene was amplified by pcr method from recombinant cloning vector pt7 - ri , which had been constructed by dr . yu xiu - ping in this laboratory and sequenced by takara biotechnology ( dalian ) co . , ltd . 于秀平博士已经构建了ri的克隆载体pt7 - ri ，测序表明与已知人胎盘ri基因的cdna序列完全相同。本文的工作是在此基础之上完成的。
Screening target ' gene of artemisinin antimalarials using drug - western from cdna expressing library : 12 b - deoxoartemisinyl - ( 4 ' - oxyacetic acid ) phenyl ether was linked to bsa by using of edc cross ! inker and the product acted as drug - probe 对重组pmd一18一t克隆载体及pqe一30表达载体双酶切，提取tctp基因和pqe一30空载体并使二者重组，然后转化m15 ，挑取阳
1 . genes of igfs , which include cds , were cloned by way of rt - pcr from the tissue of human placenta , then ligated the genes of igfs to the vector of pmd18 - t , the sequences of igfs are correct by sequencing . 2 以人胎盘组织为实验材料，采用rt - pcr方法，获得了包含cds的higf -和higf -基因片段，分别连入克隆载体pmd18 - t中，测序结果分析表明扩增的片段均为目的基因。
Reconstruction of antisense pcdnas - doc - 1r plasmid after transformation the pmeiss - fl3 - doc - 1r plasmid into e . coli cells , we have extracted the plasmid and cut doc - 1r fragment using xho i to get a 975 bp doc - 1r cdna fragment Pcdna3 - doc - 1r反义重组质粒的构建将pme18s - fl3 - doc - 1r质粒转化后进行质粒提取。用xhoi限制性内切酶从克隆载体上切取975bp的doc - 1rcdna片段。
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 。
Cloning and analyzing of the full - length cdna sequence the fragments consistent with expected length from the product of race - pcr reaction are separated , purified and recovered , after which the fragments are cloned into the vector of pmd - 1st to transform the competent cells 山na的克隆和测序将得到的3 ’一及5 ’ scepcr产物符合预期长度的片段，分离后纯化、回收，克隆到ta克隆载体pmd 18t ，转化e
Genes coding mature peptide of igfs were achieved by pcr using another pair of oligo - nucleotide primers to induce to the suitable restriction enzyme site , and the igf - i product of pcr contains 230 base pairs . igf - ii contains 219 base pairs . 3 各另外设计一对特异性pcr引物，导入适当限制性内切酶切点，以上述连有目的基因的克隆载体为模板，采用pcr方法扩增基因片段，获得长度约230bp的igf -和219bp的igf -成熟肽基因序列。
Dh5 a . methods : the target gene of the domain was obtained by rt - pcr from cultured human keratinocytes coming from isolation and culture of human skin . the target gene fragment with 647 bp was loaded into cloning vector pgem - t and transforming e . coli 方法：通过对人类皮肤组织的分离与培养，利用rt - pcr技术从培养角质形成细胞获取该功能区目的基因，并将长647bp的目的基因片段装入克隆载体pgem - t中，转化大肠杆菌dh5 ，经酶切鉴定及测序证实为目的片段后，再将目的片段装入表达载体pbv - 220转化大肠杆菌dh5进行原核表达。
Newcastle disease virus ( ndv ) strain 695 , a thermostable nature avirulent strain , were replicated in embryonated chicken eggsand its rna was extracted from allantoic fluid . referred to the reported sequence of f gene , a pair of primers were designed and synthesized . f gene of ndv b95 strain was amplified by rt - pcr , the pcr products were checked by agrose gel electrophoresis and purified by agrose gel fracion method 利用从国外引进的新城疫热稳定性天然弱毒b _ ( 95 )株接种spf鸡胚繁殖病毒，经处理后提取病毒的基因组rna ，参考国内外发表的ndv融合蛋白基因序列，设计一对特异性引物，经反转录聚合酶链式反应( rt - pcr )扩增出约1700bp大小的特异性片段，将此片段回收纯化后，利用t - a克隆技术将其克隆到pgem - t - easy克隆载体中，再转化大肠杆菌jm109感受态细胞，转化后经分子量比较、 pcr鉴定和酶切分析筛选阳性克隆。
