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  cloned
     Two novel P450 genes (CYP9A18 and CYP6AE12) from the insecticide-resistant strain (YS-FP) of Helicoverpa armigera (Hübner) were cloned with RT-PCR and RACE.
     采用RT-PCR和RACE技术克隆到2个新的棉铃虫细胞色素P450基因:CYP6AE12和CYP9A18。
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     A full-length cDNA of cab gene was cloned from the first strand of bamboo(Bambusa oldhamii) cDNA through RT-PCR and RACE methods, named as cab-BO1 (cab gene from B. oldhamii).
     采用RT-PCR技术与RACE技术,从绿竹中克隆到捕光叶绿素a/b结合蛋白基因全长cDNA,命名为cab-BO1(GenBank登记号EF061137)。
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     Designing a pair of primers according to NADL-2 strain of PPV from the Genbank,the major antigen regions of VP2 of NJ-1 strain were amplified by PCR. The PCR products were cloned into pGEM-T Easy vector. 882 bp nucleotide sequence was acquired,and 294 amino acid sequence was deduced.
     参考Gen Bank公布的NADL-2株序列,设计出1对引物,并利用该引物扩增了猪细小病毒NJ-1株VP2主要抗原基因,将其克隆到pGEM-T Easy载体上,测序获得882 bp核苷酸序列,并推导出其氨基酸序列,共编码294aa,与NADL-2株相应的序列进行比较分析,两者核苷酸同源性为99.2%,氨基酸同源性为99%;
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     The S1 gene of Infectious bronchitis virus(IBV) was segmented and cloned into the expression vector pGEX-6p-1 by gene recombination technology. The glutathiones transferase ( GST) fusion proteins were induced by IPTG.
     利用基因重组技术将鸡传染性支气管炎病毒(IBV)S1基因分段(A和B段)克隆到pGEX-6p-1载体中,用IPTG诱导表达。
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     A cDNA encoding Moschus berezovskii IFN-γ was cloned from Con-A stimulated peripheral blood mononuclear cells (PBMC) using RT-PCR and expressed in Escherichia coli.
     以RT-PCR方法,从经ConA诱导的林麝(Moschus berezovskii)外周血淋巴细胞(peripheral blood mononuclear cells,PBMC)总RNA中扩增出编码林麝IFN-γ的基因并将其克隆到pMD18-T载体上。
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  clone into
     Methods SREBP-1c and mGLUT1 cDNA clone into pSV-sport, pGL3b expression vector.
     方法采用分子克隆技术,将SREBP1c和mGLUT1cDNA分别克隆到表达载体pSVsport和pGL3b上。
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     Methods:PPARγ ,RXRα and mGLUT1 cDNA clone into pCMX and pGL3b expression vector.
     方法:采用分子克隆技术,将过氧化物酶体增殖物受体γ(PPARγ)和维甲酸类受体X(RXRα)及mGLUT1 cDNA分别克隆到表达载体pCMX和pGL3b上。
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     Methods Amplify HSP70 gene by PCR,clone into prokaryotic expression vector pET-28b,then transform to E.
     方法将PCR扩增的HSP70基因克隆到原核高效表达载体pET-28b中,转化大肠杆菌JM109。
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     Methods Extract total RNA from human liver tissue,amplify the encoding region of MBL by RT-PCR,clone into vector pGEM-T-easy,then subclone to plasmid pET-30(a) for expression under induction of IPTG.
     方法提取肝总RNA,RT-PCR扩增MBL编码区序列,并克隆到pGEM-T-easy载体上,再亚克隆到pET-30(a)上。
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     The 4 complementary DNA fragments were synthesized,and then clone into the sites of BamHⅠ and HindⅢ of the pSilencer 5.1-H1 retrovirus vector for constructing the recombinant retrovirus plasmid.
     将此双链DNA片段克隆到pSilencer 5.1-H1 Retro vector的BamHⅠ和HindⅢ位点,构建其逆转录病毒重组表达质粒。
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     OBJECTIVE To clone,express and purify a modified human plasminogen kringle 5 gene,which provides a basis for further studies on the function of RGDRGD-liteK5.METHODS The gene of RGDRGD-liteK5 was amplified from the K5 cDNA by PCR and inserted into pGEX1-λT plasmid to construct the prokaryotic fusion expression vector.
     目的改造人纤溶酶原Kringle 5(K5)基因,构建RGDRGD-liteK5融合表达质粒,并表达纯化所得RGDRGD-liteK5蛋白。 方法以人纤溶酶原K5 cDNA为模板,通过PCR得到RGDRGD-liteK5基因片段,并克隆到质粒pGEX1-λT中,构建重组原核融合表达载体pGEX-RGDRGD-liteK5;
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     ①The mB7-2 fragment obtained from plasmid pMD18-mB7-2 which was cutted by EcoRI and BamHI was inserted into vector MINV to gain vector MINV-IRES-mB7-2, which include IRES gene fragment.
     方法1将EcoRI和BamHI酶切下来的pMD18-mB7-2质粒上的mB7-2基因片段克隆到含有IRES片段的MINV质粒上,得到MINV-IRES-mB7-2载体;
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     Purpose To clon the human soluble catachol-O-Methyltransferase(COMT) into the Pet22b+ vector,expression and purification the COMT protein for researching the function of COMT.
     目的将儿茶酚胺氧位甲基转移酶基因(COMT)克隆到原核表达载体进行可溶性表达,制备纯化COMT蛋白,为深入研究COMT的功能提供材料。 方法运用分子生物学方法构建融合表达质粒Pet22b+-COMT。
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     Methods: The BXLF2 gene coding for 5'-terminal truncated of Epstein-Barr virus(EBV) gp85 was amplified from the B95-8 cell line transfected by EBV with specific primers. The PCR product was inserted into the prokaryotic expression plasmid pGEX-5T and confirmed by sequencing.
     方法:通过PCR反应从EB病毒转染的绒猴淋巴细胞系B95-8细胞中获得了gp85BXLF2基因,将此基因克隆到大肠杆菌表达载体pGEX-5T,得到阳性克隆pGEX5T-85。
短句来源
     Methods The cDNA of typeⅡ truncated receptor(ΔTβRⅡ) that lacks the cytoplasmic domain was amplified by PCR from the plasmid H2-3FF,and the eukaryotic expression plasmid pEGFP/ΔTβRⅡ was constructed by inserting the ΔTβRⅡ cDNA into the vector pEGFP-N2.The construction was introduced by transfection into L02 cells after the DNA sequence was confirmed by double digestion and sequencing.
     方法以质粒H2-3FF为模板,用PCR方法扩增得到人转化生长因子βⅡ型受体(TβRⅡ)胞外段和跨膜段的cDNA,将该cDNA克隆到真核表达载体pEGFP-N2上,构建成pEGFP/ΔTβRⅡ重组质粒。
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  cloned
A luxR homologue, designated as xooR, was cloned from X.
      
Two DREB-like genes from Bermuda grass that are induced by low-temperature or high-salt stresses were cloned using RT-PCR and RACE methods, and were named BeDREB1 and BeDREB2, respectively (GenBank accession No: AY462117 and AY462118).
      
The sequence cloned was 99% homologous to Rhodococcus erythropolis IGTS8 that was reported in the Genebank.
      
Though inter-species nuclear transfer has been introduced to construct human somatic cell cloned embryos, the effects of type, passage, and preparation method of donor cells on embryo development remain unclear.
      
In our experiment, cloned embryos were reconstructed with different passage and preparation methods of ossocartilaginous cell, skin fibroblast, and cumulus cells.
      
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  clone into
Experimental brain tumors were produced in 20 cats by stereotaxic xenotransplantation of a blastomatous glial cell clone into the internal capsule of the left hemisphere.
      
A clone bank of pRDS1 constructed by ligating fragments from aXhoII digest of a pRDS1 cosmid clone into a mobilizable plasmid was used to locate an origin of replication of pRDS1.
      
Experimental brain tumours were produced in cats by stereotactic implantation of 4 million suspended cells of a rat glioma clone into the internal capsule.
      
Injection of this clone into blastocysts generated chimeric mice that were germline competent.
      
In vivo, we used models of orthotopic implantation and liver metastasis to transplant NK4-transfected clone or mock-transfected clone into nude mice.
      
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By means of recombinant DNA technique the HBcAg gene from a cloned HBV genome was incorporated into and expressed in an E. coli strain. Using the bacterial lysate from this engineered strain we developed ELISA methods for the detection of serum anti-HBc and anti-HBc-IgM. In a parallel assay with Abbott kit of the same kind, assay results of 38 serum samples for anti-HBc (among them 23 were positive and 15 were negative according to Abbott kit) showed discrepancy only in one weakly positive sample with a relative...

By means of recombinant DNA technique the HBcAg gene from a cloned HBV genome was incorporated into and expressed in an E. coli strain. Using the bacterial lysate from this engineered strain we developed ELISA methods for the detection of serum anti-HBc and anti-HBc-IgM. In a parallel assay with Abbott kit of the same kind, assay results of 38 serum samples for anti-HBc (among them 23 were positive and 15 were negative according to Abbott kit) showed discrepancy only in one weakly positive sample with a relative concordance rate of 97.3%. Another comparative test with 40 serum samples (20 were positive and 20 were negative according to Abbott kit) showed complete agreement in the detection of anti-HBc-IgM.

采用基因工程技术,使克隆到大肠杆菌质粒上的 HBV 基因,经过体外切割重组,使其中的 HBcAg基因能在大肠杆菌中表达。以这种工程菌裂解液作为抗原,建立了检测血清中抗-HBc 和抗-HBcIgM的 ELISA方法。在与用美国Abbott厂的同类试剂盒进行的对比测定中,38份标本的抗 HBc检测结果,除 1份外,22份阳性和 15份阴性全部符合。40价标本抗HBcIgM检测结果,20份阳性和20份阴性亦完全相符。

Derivatives of transposons Tn233(CH), Tn233AE14 and Tn5 were constructed using in vitro recombination techniques. (1) Transposon Tn233 (CH) has a unique BylII restriction site, a BamHI fragment containing K88 antigen genes generated from plasmid pTK63 was ligated to this BglII site of pBK322: :Tn233 (CH) to form pBR322: :Tn233 (K88). (2) Tn233AE14 is a deletion mutant of Tn233 (CH); the deletion removed the TnpA gene of Tn233 (CH), but retained the BglII site. The same BamHI fragment was cloned into the BglII...

Derivatives of transposons Tn233(CH), Tn233AE14 and Tn5 were constructed using in vitro recombination techniques. (1) Transposon Tn233 (CH) has a unique BylII restriction site, a BamHI fragment containing K88 antigen genes generated from plasmid pTK63 was ligated to this BglII site of pBK322: :Tn233 (CH) to form pBR322: :Tn233 (K88). (2) Tn233AE14 is a deletion mutant of Tn233 (CH); the deletion removed the TnpA gene of Tn233 (CH), but retained the BglII site. The same BamHI fragment was cloned into the BglII site of pBR322: :Tn233△E14 to construct pBR322:Tn233△E14 (K88). (3) Tn5 has a unique BamHI restriction site, and pTB341 is a Tn5 inserted plasmid. Cleavage of pTB341 with BamHI yields three BamHI fragments, with fregments containing Ap' gene, IS50L and Km' gene, IS50R and Sm gene, respectively. When these fragments were re-ligated with T4-DNA ligase, a plasmid pTS40 was isolated, which also consists of these fragments, but their arrangement differs with that of pTB341. The consequence of re-ligation is that the Apr gene becomes a portion of a Tn5.The results of genetic experiments show that the K88 antigen genes and Apr gene are able to express themselves in these derivatives of transposons, and the new constructed Tn233 (K88) and Tn5 (Ap) retained their transposition ability. The Tn233Al4 (K88) is also able to transpose from pBR322: :Tn233△E14 (K88) to other plasmid in the presence of a wild type TnpA gene.

用体外重组技术构建了转座子Tn233(CH)、Tn233△E14与Tn5的3个衍生物:(1)转座子Tn 233(CH)含有单个BgBlⅡ限制位点,将从质粒pTK 63来的含K 88抗原基因的BamHI片段连接到pBR322::Tn233(CH)的BglⅡ位点上,形成了pBR322::Tn233(K88)。(2)Tn233△E14是Tn233的缺失变种,此缺失除去了Tn233(CH)上的TnpA基因,但保留了BglⅡ位点,将同上面一样的BamHI片段克隆到pBR322::Tn233△E14的BglⅡ位点上,构建了pBR322::Tn233△E14(K88)。(3)Tn5含有1个BamHI限制位点,pTB341是1个有Tn5插入的质粒,pTB341经BamHI切割后得到3个BamHI片段,每个片段分别带有Ap~r基因;IS50L与Km~r基因;IS50R与Sm~r基因,当此3个片段经T4-DNA连接酶重新连接后,分离到了1个质粒pTS40,此质粒也由此3片段组成,但它们的排列与pTB341的不同,此重新连接的结果使Ap~r基因成为Tn5中的一部分。 遗传实验结果表明,K88抗原基因与Ap~r基因在这些...

用体外重组技术构建了转座子Tn233(CH)、Tn233△E14与Tn5的3个衍生物:(1)转座子Tn 233(CH)含有单个BgBlⅡ限制位点,将从质粒pTK 63来的含K 88抗原基因的BamHI片段连接到pBR322::Tn233(CH)的BglⅡ位点上,形成了pBR322::Tn233(K88)。(2)Tn233△E14是Tn233的缺失变种,此缺失除去了Tn233(CH)上的TnpA基因,但保留了BglⅡ位点,将同上面一样的BamHI片段克隆到pBR322::Tn233△E14的BglⅡ位点上,构建了pBR322::Tn233△E14(K88)。(3)Tn5含有1个BamHI限制位点,pTB341是1个有Tn5插入的质粒,pTB341经BamHI切割后得到3个BamHI片段,每个片段分别带有Ap~r基因;IS50L与Km~r基因;IS50R与Sm~r基因,当此3个片段经T4-DNA连接酶重新连接后,分离到了1个质粒pTS40,此质粒也由此3片段组成,但它们的排列与pTB341的不同,此重新连接的结果使Ap~r基因成为Tn5中的一部分。 遗传实验结果表明,K88抗原基因与Ap~r基因在这些转座子衍生物中能够表达,而且新构建的Tn233(K88)与Tn5(Ap)仍保留着转座的能力。当在反式位置上有TnpA基因时,Tn233△E14(K88)也能从pBR322::Tn233△E14(K88)转座到其它质粒上。

Recombinant plasmids pTH3 and pTB3 were obtained by cloning the DNA fragments which contained streptomycin (Sm) resistant gene or both streptomycin and sulfonamide (Su) resistant genes from Tn233 (CH) transposon into plasmid pBR 322.By transposing Tn5 to plasmid pTH3 or pTB3,several mutants,sensitive or still resistant to Sm,have been isolated.The Tn5 insertion site were mapped by comparison of the restriction patterns of mutant plasmid DNAs with their parents plasmid DNAs.It was shown that the Sm-resistant...

Recombinant plasmids pTH3 and pTB3 were obtained by cloning the DNA fragments which contained streptomycin (Sm) resistant gene or both streptomycin and sulfonamide (Su) resistant genes from Tn233 (CH) transposon into plasmid pBR 322.By transposing Tn5 to plasmid pTH3 or pTB3,several mutants,sensitive or still resistant to Sm,have been isolated.The Tn5 insertion site were mapped by comparison of the restriction patterns of mutant plasmid DNAs with their parents plasmid DNAs.It was shown that the Sm-resistant gene carried by pTH3 directed the synthesis of a po-lypeptide with molecular weight of about 32K in Escherichia coli maxicell.Based upon these data,the location and length of Sm-resistant gene on the pTH3 DNA were mapped With the similar method,the location and length of Su resistant gene on the pTB3 DNA were also mapped.With the polar effect of Tn5,it is supposed that Sm-and Su-resistant genes do not compose a single opron in the Tn233 (CH) transposon DNA.

重组质粒pTH3和pTB3分别是转座子Tn233(CH)中含Sm抗性基因和同时含Sm和Su抗性基因的DNA片段克隆到pBR322上得到的。将Tn5转座到pTH3或pTB3上,分离到一些对Sm敏感或仍有抗性的突变株。比较变种及亲本质粒DNA的限制图谱,测出了Tn5的插入位点,并测得pTH3的Sm抗性基因在大肠杆菌极大细胞中指令一个分子量约为32K的多肽合成。据此我们绘制了pTH3中Sm抗性基因的位置与长度。用相似方法也绘制出了Su抗性基因在pTB3上的位置与长度。利用Tn5的极性效应,推测Tn233(CH)中Su和Sm抗性基因不构成一个操纵子。

 
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