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In this experiment, His 5 gene was inserted into Ura 3 gene and constructed a new plasmid pLRH33, which contained a complete His 5 gene and partial Ura 3 gene at the ends of His 5 gene.

实验将His 5基因插入到一个Ura 3基因的中间，构建了一个新的质粒pLRH33，从而打断了Ura 3 基因使之不能表达。

Methods: The coding region of superoxide dismutase was amplified using PCR method from the E.co1i genome. The PCR product was cloned into PUC19-T vector and sequenced. In addition, the cloned coding region of Mn-SOD was inserted into the expression vector PET-28a to form the recombinant plasmid PET-28a-Mn-SOD and was then transformed into E.coli BL21 for expression.

用PCR方法从大肠杆菌2号基因组中扩增Mn-SOD基因编码区，克隆到pUCm-T Vector，测定核苷酸序列；再将基因编码区克隆到原核表达载体PET-28a，构建含Mn-SOD基因的重组表达质粒，转化到大肠杆菌BL21中进行IPTG诱导表达。

objective: to achieve the soluble expression of mn-sod gene in e.coli and assay the enzyme activity of the expressed product. methods: the coding region of superoxide dismutase was amplified using pcr method from the e.coli genome. the pcr product was cloned into puc19-t vector and sequenced.in addition,the cloned coding region of mn-sod was inserted into the expression vector pet-28a to form the recombinant plasmid pet-28a-mn-sod and was then transformed into e.coli bl21 for expression.

超氧化物歧化酶(superoxide dismutase，sod)是细胞体内歧化超氧阴离子自由基(o-2)的一个抗氧化酶，按其结合的金属性离子根据其中金属辅基的不同可分为4类：cu/zn-sod、mn-sod、fe-sod和ni-sod，它们通过催化超氧阴离子自由基0-2发生歧化反应，达到清除o-2的效果，具有防御氧毒性、增强机体抗辐射损伤能力、防衰老以及治疗某些肿瘤、炎症、自身免疫疾病等功效，广受国内外科研工作者的关注和重视[1]。

The percentage of plasmid in vibrio cholerae from different countries was reviewed and the biological characteristics were introduced .

本文对国外报道的霍乱弧菌中的质粒携带率以及质粒决定的生物学功能进行综述。

The plasmid of pEGFP-N1 was preserved in our laboratory.②The rat vibrissa follicles were dissected under a stereomicroscope. The dermal sheath was moved by incubated in dispase. The bulge regions of the hair follicles in anagen phase were carefully cut between the arrector pili muscle and the sebaceous gland, and then incubated with a mixture of trypsin and EDTA. The cell suspension was selected, and cultured in 10% fetal bovine serum DMEM/F12 FAD medium. After 7 days culture, HFSC was obtained by rapid adhering on collagen Ⅳ.

实验方法：大鼠在体视显微镜下分离出含真皮鞘的完整毛囊，dispase消化，将毛囊从真皮鞘中挤出，收集形态完好且处于生长期的毛囊，分别在毛球部上端、皮脂腺下端横切毛囊，取中间部分置于胰酶和EDTA中联合消化，向所得细胞悬液中添加含10%胎牛血清DMEM/F12完全FAD培养基，常规培养7 d后，采用IV型胶原快速贴壁法两次筛选以分离纯化大鼠毛囊干细胞。

Recombinant plasmid pSVH 7 DNA of avian influenza virus H7 subtype heamagglutinin gene was encapsulated with DC-chol/DOPE liposomes and PC/chol/SA liposomes separately. Two-week old SPF chickens were intramuscularly inoculated with 50 μ g/0.2ml of the liposome entrapped PSVH 7 DNA. Four-weeks later, each chicken was challenged with 0.1ml 〓 AIV . One week after the challenge, the secretion of the cloacas was collected and transfected to chicken embryos to isolate the virus. The virus was isolated from 6/6 of the control group, 1/6 of the naked DNA group, 1/6 of the PC/chol/SA entrapped DNA group and 0/6 of the DC-chol/DOPE liposome entrapped group. The HI antibody titers (log2) of the four groups were 6. 83±0.98, 7. 0±1. 26, 7. 83±1. 17 and 8. 00±0.89 respectively 1-week after challenge, and 8. 5±0.55, 8. 17±0.82, 8. 68±0.45 and 9. 33±0.54 respectively 2-week after challenge. The results showed that inoculation of liposome entrapped DNA significantly enhanced resistance to virosis in animals.

将含禽流感病毒H7亚型血凝素基因的重组质粒pSVH7用DC-chol阳离子脂质体和胆固醇／卵磷脂／十八胺脂质体包裹，免疫2周龄SPF鸡，4周后用同型禽流感病毒进行人工感染，1周后采集泄殖腔分泌物分离病毒，结果未免疫组6／6分离到病毒，裸质粒DNA免疫组1／6分离到病毒胆固醇／卵磷脂／十八胺脂质体包裹DNA免疫组1／6分离到病毒，DC-chol脂质体DNA免疫组没有分离到病毒（0／6）：人工感染后1周各组的HI抗体（Log2）分别为6.38±0.98，7.00±1.26，7.83±1.17，8.00±0.89，2周后为8.50±0.55，8.67±0.82，8.68±0.45，9.33±0.52，脂质体包裹组在同期均高于未免疫组和裸DNA免疫组，表明脂质体包裹质粒DNA免疫动物后，能增加动物对病毒感染的抵抗力和反应能力。

HRP was coupled with the fluorochrome FITC and encapsulated with liposome . pEGFP-N1 plasmid (expressing green fluorescent protein) with encapsulation in commercially available liposome was prepared. The eyes were enucleated 1, 4 and 8 weeks after zonule rupture and anterior segments comprising lenses were incubated in medium containing one of these components. Cryo-sections were made and translocation of fluorescent macromolecule from the medium into the lens and green fluorescent protein expression in the epithelium were observed by fluorescent microscopy.

制备FITC标记的HRP脂质体；制备pEGFP-N1质粒（表达绿色荧光蛋白的质粒），并用脂质体包裹；豚鼠悬韧带部分离断后1周、4周、8周取含晶状体的眼前段标本分别在含有FITC-HRP复合物、pEGFP-N1质粒的培养基中孵育，冰冻切片，荧光显微镜观察FITC-HRP复合物进入晶状体和pEGFP-N1质粒在晶状体内表达的情况，比较悬韧带离断侧与非离断侧的差异。3。

Abjective to express the Hepatitis A virus genome cDNA attenuated salmonella Typhimurium and to find the immunity to hepatitis A virus after administrated that salmonella Typhimurium with hepatitis A virus gemome to the mousehepatitis A virus coding region cDNA were inserted expressive plasmid PBV221 and then were transform into E coli DH5α.

使甲型肝炎病毒基因组cDNA能够在减毒伤寒杆菌中表达，从而通过口服此种减毒伤寒杆菌达到对甲肝免疫的作用。将甲肝病毒编码区cDNA插入高效表达质粒PBV221，转化大肠杆菌DH5α，筛选并鉴定阳性克隆。

BALB/c mice were immunized with plasmid TR421-hCGβ or mock DNA three times. Two weeks after immunization, spleen cells from the immunized mice were harvested, and then analyze the CTL activity against Sp2/0-ehCGβ cells. The splenocytes derived from the immunized mice were adoptively transferred to the normal mice, which were subsequently given injections i.

以TR421-hCGβ质粒实施基因免疫，免疫后检测小鼠脾细胞，特异性细胞毒活性；同期将脾细胞过继免疫给正常小鼠，以过继TR421-hCGβ质粒免疫小鼠脾细胞为实验组、过继TR421质粒免疫小鼠脾细胞为对照组，检测脾细胞杀伤效应。

It was identified that the nif gene for aerogenic P. agglomerans strain waslocated on plasmid while for anaerogenic strains were located on chromosomes usingKlebsiella pneumoniae nifHDK as probe.

用nifHDK为探针进行Southern杂交证明固氮成团泛菌产气型的菌株固氮基因定位在质粒上，而不产气型菌株固氮基因定位在染色体上。

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！