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PCR productsdigested by SnaB Ⅰ and Not Ⅰ were inserted into the correspondingrestriction site in the expression plasmid pPIC9K.

用限制性内切酶 SnaBⅠ和 NotⅠ酶切 NGF cDNA 的 PCR 产物，将其插入到载体 pPIC9K 中相应的限制性内切酶位点，构建表达载体pPIC9K-NGF.3。

Methods Digest recombinant plasmid pU57-SOD with SnaB Ⅰ and Not Ⅰ and insert the obtained SOD gene into expression vector pPIC9k.

方法根据酵母密码子偏好性，合成人Cu，Zn-SOD基因，克隆至真核表达载体pPIC9k中，转化毕赤酵母GS115。

Several fragments containing the replication origin were cloned into replicon probe vector pUC-MEV constructed in this study. After screening a 4. 8kb PvuⅡ-SnaBⅠ fragment were determined to be the minimum replication region of the plasmid.

将包含复制起始原点在内的数个片段分别插入构建的嗜盐古生菌的复制子探针载体pUC-MEV中，通过筛选得到了pHH205最小复制功能片段。

A complete bovine 18ku-bFGF gene was cloned by nested-PCR and subcloned into expression vector pET-28a.The recombinant plasmid pET-28a-bFGF was transformed into Escherichia coli BL21.At 25℃,recombinant protein was induced by 0.5mmol/L IPTG for 5 hours.Supernatant of cell lysate was purified using Ni-NTA method and the products was submitted to detect the bioactivity by Western-blotting,which indicated that the fusion protein contained recombinant bovine bFGF.Results showed that recombinant bovine bFGF was produced and solubly existed in the supernatant.NIH-3T3 fibroblasts were used to detect the bioactivity of the fusion protein.

采用巢式PCR方法克隆了牛18ku-bFGF基因完整的编码序列，并构建了原核表达载体pET-28a-bFGF，将其转化大肠杆菌BL21，在25℃低温条件下，用0.5mmol/L IPTG诱导表达5h，用Ni-NTA亲和纯化细胞裂解上清液，经Western-blotting检测，结果显示，在特定的诱导条件下，重组牛bFGF基因在大肠杆菌中获得了表达，并且主要以可溶性状态存在于细胞中。

The open reading frame of Spinacia oleracea Betaine Aldehyde Dehydrogenase was retrieved from Spinacia oleracea and inserted into the Agrobacterium tumefaciens binary vector pBin438, which was driven by CaMV35S promoter, and produced the new binary vector pBSB. A. tumefaciens LBA4404 carrying this plasmid was used in genetic transformation of plants. Forty-five primary transgenic plants were detected by PCR and verified by the Southern blotting from 65 regenerated plants, of which 27 transgenic plants had only one copy of T-DNA.

摘 要：分离出菠菜甜菜碱醛脱氢酶基因构建成由CaMV35S驱动的双元植物表达载体pBSB，农杆菌菌株LBA4404携带该载体转化棉花，获得转基因棉花植株。65株转基因植株经过PCR筛选、Southern blotting分析证明有45株为成功的转化株，外源基因已经被整合到棉花的染色体组中，并以单拷贝插入居多。

The calli, derived from immature embryos, of the supersweet corn inbred line was transformed with the plasmid DNA containing Bt gene via the particle bombardment-mediated method. Through optimizing regeneration and rooting conditions in supersweet corn immature embryo culture, a stable, highly efficient plant post-transformation regenerating system has been established.

以超甜玉米幼胚为愈伤组织诱导外植体，利用基因枪法将苏云金杆菌杀虫蛋白基因转化胚性愈伤组织，通过优化再生和生根培养条件建立了稳定、高效的超甜玉米转基因再生成株体系。

Full-length plasmid DNA was amplified with the above primers with a high-fidelity thermostable DNA polymerase capable of long-rang amplification.

用能扩增长片段的高保真耐热 DNA聚合酶扩增全长的质粒DNA，直接转化大肠杆菌。

We prepared critical size segmental bone defects in rat tibiae and transplanted GAM consisting of boine atelocollagen and expression plasmid ector (bmp2), which encodes human BMP2, with or without CaP.

我们在大鼠胫骨上做了临界大小的节段性骨缺损，并移植上包含牛次发育胶原、表达bmp2质粒的GAM，后者编码人BMP2，不管有没有磷酸钙沉淀物的存在。

An in vitro transcription recombinant plasmid 5T1d was obtained after subcloning the target gene of clone 5T1d,which contained mutant HCV 5 non-coding region cDNA,into a transcriptive vector pCDNA Ⅰ.

采用含HCV5非编码区(5NCR)缺失突变的重组质粒5T1d，将其目的基因亚克隆到体外转录载体pCDNAⅠ多克隆位点上，获得转录重组体5T1d。

The gene of transglutaminase was amplified from the chromosome of Bacillus subtilis DB403 by PCR and translationally fused with thioredoxin gene in the plasmid pET32a to achieve the expression vector pET32-BTGase.

利用PCR技术，从枯草杆菌DB403染色体上扩增出谷氨酰胺转胺酶基因，将其克隆到大肠杆菌载体pET32a中，成功构建谷氨酰胺转胺酶表达载体pET32-BTGase，并转化大肠杆菌BL21 (DE3)。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！