Existence and Functions of Neurotens in Human Early Placental Villi

ＥｘｉｓｔｅｎｃｅａｎｄＦｕｎｃｔｉｏｎｓｏｆＮｅｕｒｏｔｅｎｓｉｎＨｕｍａｎＥａｒｌｙＰｌａｃｅｎｔａｌＶｉｌｌｉＺＨＡＮＧＣｈｏｎｇ－ｌｉ（张崇理）;ＣＨＥＮＧＬｉ－ｒｅｎ（程丽仁）,ＳＨＥＮＷｅｉ－ｂｉｎ（沈卫斌）;ＹＩＮＨｏｎｇ（殷红）;ＨＵ...     

Expression of Porcine Growth Hormone Gene in CHO Cell

ＥｘｐｒｅｓｓｉｏｎｏｆＰｏｒｃｉｎｅＧｒｏｗｔｈＨｏｒｍｏｎｅＧｅｎｅｉｎＣＨＯＣｅｌｌＣＨＥＮＱｉｎｇ－ｘｕａｎ（陈清轩）;ＨＥＸｉｎ（何新）;ＤＥＮＧＨｕｉ－ｎａｎ（邓辉南）（ＩｎｓｔｉｔｕｔｅｏｆＤｅｖｅｌｏｐｍｅｎｔａｌＢｉｏｌｏｇｙ,Ａｃ...     

Factors Affecting Electro－Fusion and Electro－Activation In Serial Nuclear Transplantation In Goat（Carpa hircus） Embryo

ＦａｃｔｏｒｓＡｆｆｅｃｔｉｎｇＥｌｅｃｔｒｏ－ＦｕｓｉｏｎａｎｄＥｌｅｃｔｒｏ－ＡｃｔｉｖａｔｉｏｎＩｎＳｅｒｉａｌＮｕｃｌｅａｒＴｒａｎｓｐｌａｎｔａｔｉｏｎＩｎＧｏａｔ（Ｃａｒｐａｈｉｒｃｕｓ）ＥｍｂｒｙｏＷＡＮＧＹｕ－ｇｅ（王玉阁）;ＺＯＵＸ...     

Inhibition of Acrosomal Enzymes by Gossypol Is Related to Its Antifertility Action

ＩｎｈｉｂｉｔｉｏｎｏｆＡｃｒｏｓｏｍａｌＥｎｚｙｍｅｓｂｙＧｏｓｓｙｐｏｌＩｓＲｅｌａｔｅｄｔｏＩｔｓＡｎｔｉｆｅｒｔｉｌｉｔｙＡｃｔｉｏｎＹＵＡＮＹｕ－ｙｉｎｇ;（袁玉英）ＺＨＡＮＧＹａｎ－ｌｉｎ;（张燕林）ＳＨＩＱｉ－ｘｉａｎ（石其贤）（Ｚｈｅ...     

Effect of Plant AntifreezeProteins on Porcine Embryo to Cryopreservaton

ＥｆｆｅｃｔｏｆＰｌａｎｔＡｎｔｉｆｒｅｅｚｅＰｒｏｔｅｉｎｓｏｎＰｏｒｃｉｎｅＥｍｂｒｙｏｔｏＣｒｙｏｐｒｅｓｅｒｖａｔｏｎ￥ＦＥＩＹｕｎ－ｂｉａｏ（费云标）;ＷＥＩＬｉｎｇ－ｂｏ（魏令波）;ＧＡＯｓｕ－ｑｉｎ（高素琴）;ＺＨＡＯＳｈｕ－ｈｕｉ（赵...     

MPF Induction of Microtubule Assembly at Interphase Kinetochore of CHO Cells

ＭＰＦＩｎｄｕｃｔｉｏｎｏｆＭｉｃｒｏｔｕｂｕｌｅＡｓｓｅｍｂｌｙａｔＩｎｔｅｒｐｈａｓｅＫｉｎｅｔｏｃｈｏｒｅｏｆＣＨＯＣｅｌｌｓＦＥＮＧＭｅｉ;（冯梅）ＺＨＡＮＧＨｕａｎ－ｘｉａｎｇ;（张焕相）ＷＡＮＧＹｏｎｇ－ｃｈａｏ;（王永潮）ＷＡＮＧＹｕｅ...     

生态交错带（Ecotone）理论研究进展

生态交错带（Ｅｃｏｔｏｎｅ）理论研究进展高洪文（东北师范大学草地研究所,长春１３００２４）ＡｄｖａｎｃｅｍｅｎｔｏｆＴｂｅｏｒｅｔｉｃａｌＲｅｓｅａｒｃｈｉｎＥｃｏｔｏｎｅ．￥ＧａｏＨｏｎｇｗｅｎ（ＩｎｓｔｉｔｕｔｅｏｆＧｒａｓｓｌａｎｄＲｅ－ｓｅａ...     

Detection and Analysis of an Estrous－associated Oviductal Glycoprotein DNA Fragment from Primates by PCR

ＤｅｔｅｃｔｉｏｎａｎｄＡｎａｌｙｓｉｓｏｆａｎＥｓｔｒｏｕｓ－ａｓｓｏｃｉａｔｅｄＯｖｉｄｕｃｔａｌＧｌｙｃｏｐｒｏｔｅｉｎＤＮＡＦｒａｇｍｅｎｔｆｒｏｍＰｒｉｍａｔｅｓｂｙＰＣＲ￥ＣＨＥＮＱｉｎｇ－ｘｕａｎ（陈清轩）;ＣｌａｙｔｏｎＥ．Ｗａｌｔｏ...     

A Protein Extracted from Mouse Sperm That Plays an Important Role in Fertilization

ＡＰｒｏｔｅｉｎＥｘｔｒａｃｔｅｄｆｒｏｍＭｏｕｓｅＳｐｅｒｍＴｈａｔＰｌａｙｓａｎＩｍｐｏｒｔａｎｔＲｏｌｅｉｎＦｅｒｔｉｌｉｚａｔｉｏｎＺＨＵＡＮＧＤａ－ｚｈｏｎｇ;（庄大中）,ＺＨＡＮＧＴｉａｎ－ｙｉｎ（张天荫）,ＣＨＥＮＤａ－ｙｕａｎ;（陈大...     

Incorporation of Resistance Gene into Tomato using Binary Vector System

ＩｎｃｏｒｐｏｒａｔｉｏｎｏｆＲｅｓｉｓｔａｎｃｅＧｅｎｅｉｎｔｏＴｏｍａｔｏｕｓｉｎｇＢｉｎａｒｙＶｅｃｔｏｒ　ＳｙｓｔｅｍＺＨＡＯＳｈｕ－ｈｕｉ（赵淑慧）;ＸＵＥＧｕｏ－ｘｉｏｎｇ（薛国雄）（ＩｎｓｔｉｔｕｔｅｏｆＤｅｖｅｌｏｐｍｅｎｔａｌＢｉｏ...     

Construction of Plant Expression Vector with Double Resistance to Virus and Insect and Identification of Transformation in Tomato

By in situ hybridization of bacterium clone and analysis of restriction enzyme digestion, both CMV-cp gene and Bt-toxin gene were inserted one by one into T-DNA of binary plant expression vector pea. The reconstructed plasmid was named pE14. Then, tomato was transformed with pE14 mediated by Agrobacterium tumefaciens GV311-SE, four regenerated tomato plants were obtained on the MS medium containing 100 μg/mL kanamycin. Assay of nopaline, dot blotting of tomato genomic DNA and PCR amplication of CMV-cp gene and Bt-toxin gene from genomic DNA showed that CMV-cp gene and Bt-toxin gene were transferred into the four regenerated tomato plants simultaneously with T-DNA, and no recombination of genes occurred. RNA dot blotting showed that two of them could express simultaneously the CMV-cp gene and Bt-toxin gene proteins. The resistances to virus and insect of the transgenic tomato plants will be tested in their F1 and F2 regenerations.     

利用双T-DNA载体系统培育无选择标记转基因烟草

建立了一种利用双T-DNA载体培育无选择标记转基因植物的方法.通过体外重组构建了双T-DNA双元载体pDLBRBbarm.载体中,选择标记nptⅡ基因和另一代表外源基因的bar基因分别位于2个独立的T-DNA.利用农杆菌介导转化烟草(Nicotiana tabacum L.),在获得的转化植株中,同时整合有nptⅡ基因和bar基因的频率为59.2%.对4个同时整合有nptⅡ和bar基因植株自交获得的T1代株系进行检测分析,发现在3个T1代株系2个T-DNA可以发生分离,其中约19.5%的转基因T1代植株中只存在bar基因而不带选择标记nptⅡ.这一结果说明双T-DNA载体系统能有效地用于培育无选择标记的转基因植物.研究还利用位于2个不同载体上的nptⅡ基因与 bar基因通过农杆菌介导共转化烟草,获得共转化植株的频率为20.0%～47.4%,低于使用双T-DNA转化的共转化频率.     

A set of novel Ti plasmid-derived vectors for the production of transgenic plants

Summary We describe in this paper the construction and use of a set of novel Ti plasmid-derived vectors that can be used to produce transgenic plants. These vectors are based on one of two strategies: 1) double recombination into the wild-type Ti plasmid of genetic information flanked by two T-DNA fragments on a wide-host range plasmid; 2) the binary vector strategy. The vector based on the double recombination principle contains a kanamycin resistance gene for use as a plant selectable marker, a polylinker for the insertion of foreign genes, and a nopaline synthase gene. The vector was constructed such that a disarmed T-DNA results from the double recombination event. The binary vector combines several advantageous features including an origin of replication that is stable in Agrobacterium in the absence of selection, six unique sites for insertion of foreign genes, an intact nopaline synthase gene, and a kanamycin resistance marker for selection of transformed plant cells. All of these vectors have been used to produce tobacco plants transformed with a variety of foreign genes.     

Green fluorescent protein as a vital elimination marker to easily screen marker-free transgenic progeny derived from plants co-transformed with a double T-DNA binary vector system

We investigated the potential of a novel double T-DNA vector for generating marker-free transgenic plants. Co-transformation methods using a double T-DNA vector or using mixture of two Agrobacterium tumefaciens strains were compared, and showed that the double T-DNA vector method could produce marker-free transgenic tobacco (Nicotiana tabacum L.) plants more efficiently. A dual marker double T-DNA vector was then constructed by assembling the green fluorescent protein (GFP) gene mgfp5 and the neomycin phosphotransferase gene nptII into the same T-DNA. The frequency of co-transformants produced by this vector was 56.3%. Co-expression of mgfp5 and nptII was found in 28 out of 29 T₁ lines, and segregation of the reporter -glucuronidase gene, gusA, from mgfp5 to nptII was found in 12 out of 29 T₁ lines. Therefore, GFP could be used as a vital marker to improve the transformation efficiency and to easily monitor the segregation of marker genes, thus facilitating screening of marker-free progeny.     

Recovery of transgenic rice plants expressing the rice dwarf virus outer coat protein gene (S8)

 The coding region of the eighth largest segment (S8) of the rice dwarf virus (RDV) was obtained from a RDV Fujian isolate. It was then cloned into pTrcHisA for expression in E. coli and into vector pE3 for plant transformation. By using callus derived from mature rice embryos as the target tissue, we obtained regenerated rice plants after bombardment of the former with plasmid pE3R8 containing the RDV S8 gene and the marker gene neomycin phosphotransferase (NPT II). Southern blotting confirmed the integration of the RDV S8 gene into the rice genome. The expression of the outer coat protein in both E. coli and rice plants was confirmed by western blotting. The recovery of transgenic rice plants expressing S8 gene is an important step towards studying the function of the RDV genes and obtaining RDV-resistant rice plants. Received: 1 March 1996 / Accepted: 2 August 1996     

The proteolytic cleavage of PE2 to envelope glycoprotein E2 is not strictly required for the maturation of Sindbis virus.

The ionophore monensin has been shown previously to block the maturation of Sindbis virus as well as prevent the cleavage of pE2 to E2 when applied to cells in high concentration. We found that a moderate dose of monensin reduced virus titer and inhibited the cleavage of pE2 to E2. Under these conditions, pE2 appeared on the cell surface in a form susceptible to lactoperoxidase-mediated iodination. This pE2 was incorporated into virions, replacing E2. PE2-containing virions had a normal PFU-to-particle ratio, cosedimented with normal virus, and retained a normal morphology when negatively stained preparations were examined by electron microscopy. We conclude that the cleavage of pE2 to form E2 is not an absolute prerequisite for virus maturation. Recently, Russell et al. have reached a similar conclusion (D. L. Russell, J. M. Dalrymple, and R. E. Johnston, J. Virol. 63:1619-1629, 1989).     

转基因水稻T—DNA侧翼序列的扩增与分析

利用现有的转抗白叶枯病基因Xa21的水稻材料,通过TAIL－PCR技术扩增出携带Xa21基因的T－DNA的侧翼序列,对24个有效扩增片段的序列分析结果表明,其中14个侧翼序列是水稻DNA,9个含载体主干序列,1个是外源基因Xa21片段,14个T－DNA侧翼的水稻DNA序列与直接转化法外源基因整合位点的基因组序列具有不同的特点,这些T－DNA在水稻染色体上整合后其两端序列的特点类似于在转基因双子叶植物中观察到的现象,在含主干序列的侧翼序列（37．5％,9／24）,中,载体主干序列是以不同的类型出现的。     

Use of targeted insertional mutagenesis to determine whether chondroitin lyase II is essential for chondroitin sulfate utilization by Bacteroides thetaiotaomicron.

Bacteroides thetaiotaomicron produces two inducible chondroitin lyases (I and II) when it is grown on chondroitin sulfate. Both enzymes have very similar biochemical properties. To determine whether both enzymes are required for growth on chondroitin sulfate, we constructed a Bacteroides suicide vector, pE3-1, and used it to create an insertional mutation that interrupts the chondroitin lyase II gene of Bacteroides thetaiotaomicron. pE3-1 contains a 4.4-kilobase cryptic B. eggerthii plasmid (pB8-51), the Escherichia coli cloning vector pBR328, and the EcoRI D fragment from the conjugative B. fragilis plasmid pBF4. A 0.8-kilobase fragment from the center of the B. thetaiotaomicron chondroitin lyase II gene was inserted in pE3-1 to create pEG817. Although, pEG817 is stably maintained in E. coli and can be mobilized into B. thetaiotaomicron by the IncP plasmid R751, pEG817 is not maintained as a plasmid in Bacteroides spp. When pEG817 was mobilized into B. thetaiotaomicron, with selection for a drug marker on pEG817, transconjugants were obtained which had pEG817 inserted into the chondroitin lyase II gene. Western blot analysis was used to confirm that intact chondroitin lyase II is not produced in the mutant. The mutant was able to utilize chondroitin sulfate as a sole source of carbon, although no active chondroitin lyase II was produced. Thus chondroitin lyase I alone appears to be sufficient for growth on chondroitin sulfate. The mutant also had some minor changes in its outer membrane protein profile. However, there was no evidence that any of the major chondroitin sulfate-associated polypeptides in the outer membrane were affected by the insertion in the chondroitin lyase II gene.     

无选择标记和载体骨干序列的Xa21转基因水稻的获得

利用双右边界T-DNA载体通过根癌农杆菌介导法将水稻白叶枯病广谱抗性基因Xa21导入杂交稻重要恢复系C418中。T0代共获得27个独立转基因株系,通过田间抗性鉴定与PCR分析,有17个株系的Xa21基因分子鉴定为阳性,且对白叶枯病原菌P6生理小种具有抗性。通过对17个株系的后代植株进行田间抗性鉴定,分子标记辅助选择及Southern杂交分析,结果显示4个株系的T1代植株中能分离出无潮霉素标记基因的Xa21转基因植株。无选择标记Xa21转基因株系的获得率为15%。PCR检测还表明,这些无选择标记的Xa21转基因植株不带有载体骨架序列。通过对转基因后代进一步的抗性鉴定与PCR辅助选择,获得了无选择标记和载体骨架序列的转基因Xa21纯合的抗白叶枯病水稻。