The present study aimed at the preparation of monoclonal antibody against the recombinant PthA-NLS and the isolation of the relative ScFv （single chain variable fragment） genes, providing the possibility to better u...The present study aimed at the preparation of monoclonal antibody against the recombinant PthA-NLS and the isolation of the relative ScFv （single chain variable fragment） genes, providing the possibility to better understand the pathogenesis mechanism via PthA, and developing proper construct for future experimentation to obtain citrus plants resistant to canker disease by transformation and plant antibody techniques. The recombinant polypeptide PthA-NLS was injected into Balb/c mice to produce monoclonal antibody. Total RNA was isolated from the hybridoma cell line 3D10H2 which secreted anti- PthA-NLS McAb, and the variable region genes were amplified with specific primers by RT-PCR and SOE-PCR （splicing by overlap extension）, and then the ScFv gene was isolated. The recombinant ScFv gene was cloned into pGEM-T and pET32a（＋） vector. The later plasmid was transferred into E. coli BL21 （DE3） and the expression of the recombinant protein was induced. Three cell lines producing monoclonal antibody against PthA-NLS were acquired and named 1C8H1, 2D12B6, and 3D8A10. The recombinant ScFv gene of about 750 bp was constructed. The sequencing results showed that the ScFv gene consists of a 360 bp heavy chain, a 342 bp light chain, and a 45 bp linker region. The recombinant fusion ScFv protein was expressed by IPTG induction, and a 44.5 kDa of recombinant fusion protein was obtained. In conclusion, we obtained three cell lines stably producing monoclonal antibody specifically bound to PthA-NLS, and the relative ScFv gene was constructed and successfully expressed in E. coli. These results may play an important role in further understanding the pathogenesis mechanism and in the development of possible citrus resistant to canker disease by genetic transformation and plant antibiobody.展开更多
Total RNA was isolated from the phloem of young shoots and retro-transcripted to cDNA by reverse transcriptasepolymerase chain reaction （RT-PCR） with specific primers; the amplified cDNA fragments were sequenced for...Total RNA was isolated from the phloem of young shoots and retro-transcripted to cDNA by reverse transcriptasepolymerase chain reaction （RT-PCR） with specific primers; the amplified cDNA fragments were sequenced for identification of subtypes. In this study, an effective RT-PCR technique was established for detecting citrus viroids, and three citrus viroids were recognized, namely, CEVd, CVd-II, and CVd-III. The latter two were identified for the first time in China. The sequences of CVd-Ⅱ were different from all the three subtypes reported in the GenBank and seemed to be a new subtype. CVd-Ⅲb detected in the present study could be further developed as a dwarfing agent.展开更多
A new technology for harnessing the dye polluted water and dye collection was developed. It is based on the enhanced evaporation by using solar, wind and air temperature energy and additional heat-electric energy. It ...A new technology for harnessing the dye polluted water and dye collection was developed. It is based on the enhanced evaporation by using solar, wind and air temperature energy and additional heat-electric energy. It consists of four parts: (1) evaporation carrier system (evaporation carrier and frame for evaporation carrier) for polluted water; (2)polluted water circulating system (pumping-spraying-collecting); (3)heating system; (4)workshop with polluted water reservoir-tanks and rainfall prevention roof. The polluted water was (heated in case necessary) sprayed to the evaporation carrier system and the water was evaporated when it moved in the space and downward along the carrier mainly by using natural (solar, wind and air temperature energy). In case, when there is no roof for the carrier system, the polluted water can be stored in the reservoirs (storage volume for about 20 days). The first 10-25 mm rainfall also need to be stored in the reservoirs to meet the state standard or discharging wastewater. The dye may be collected at the surface in the reservoir-tanks and the crystallized salt may be collected at the bottom plate. The black-color wastewater released by the factory is no more discharged to the surface water system of Taihu Lake Basin. About 2 kg dye and 200 kg industrial salt may be collected from each tone of the polluted water. The non-pollution production of dye may be realized by using this technology with environmental, economical and social benefits.展开更多
The title compound (Ⅰ) has been synthesized by the reaction of [RuCl2(PPh3)3], closo-[B10H10]2- and PhCH(OH)COOH in (CH3)2CHOH solution. The re-action gave two main products Ⅰ and Ⅱ. Crystals of the title compound ...The title compound (Ⅰ) has been synthesized by the reaction of [RuCl2(PPh3)3], closo-[B10H10]2- and PhCH(OH)COOH in (CH3)2CHOH solution. The re-action gave two main products Ⅰ and Ⅱ. Crystals of the title compound (Ⅰ) suitable fora single crystal X-ray diffraction analysis were recrystailized from n-pentane/CH2Cl2 so-lution. Its structure was determined by X-ray crystallography analysis. The crystal of{(PPh3)2RuB10 H8 [(CH3)2CHO]2} (Ⅰ) is monoclinic, space group P21/n, Mr =860. 01, with a= 12. 927(2), b= 23. 672(6), c= 14. 090(4) A, β=93. 22(2)°, V=4305 A3, Dc= 1. 327 g/cm3, Z= 4, λ(MoKα) =0. 71073A, μ=4. 6 cm-1, F(000) =1776, R = 0. 059, Rw = 0. 075, S = 2. 62. The cluster structure is based on a closoeleven-vertex {RuB10 } polyhedron skeleton with two replacing isopropoxy and twoPPh3 groups.展开更多
文摘采用乙醇和二氯甲烷分别浸提核桃叶片,获得粗提物,并在室内测定了不同浓度粗提物溶液对马铃薯块茎蛾(Phthorimaea operculella)成虫的产卵选择性及幼虫钻蛀性的影响。结果表明:两种核桃叶粗提物均对马铃薯块茎蛾的产卵以及幼虫钻蛀表现出强烈的抑制效果,而且这种抑制效果随着粗提物浓度的上升而增强;2~162 g E/L的两种粗提物溶液均不影响马铃薯块茎蛾雌蛾的交配行为;两种粗提物对块茎蛾幼虫钻蛀行为的药效能够持续36 h。
基金supported by the Scientific and Technical Project of Hunan Province,China(04NK1005)
文摘The present study aimed at the preparation of monoclonal antibody against the recombinant PthA-NLS and the isolation of the relative ScFv （single chain variable fragment） genes, providing the possibility to better understand the pathogenesis mechanism via PthA, and developing proper construct for future experimentation to obtain citrus plants resistant to canker disease by transformation and plant antibody techniques. The recombinant polypeptide PthA-NLS was injected into Balb/c mice to produce monoclonal antibody. Total RNA was isolated from the hybridoma cell line 3D10H2 which secreted anti- PthA-NLS McAb, and the variable region genes were amplified with specific primers by RT-PCR and SOE-PCR （splicing by overlap extension）, and then the ScFv gene was isolated. The recombinant ScFv gene was cloned into pGEM-T and pET32a（＋） vector. The later plasmid was transferred into E. coli BL21 （DE3） and the expression of the recombinant protein was induced. Three cell lines producing monoclonal antibody against PthA-NLS were acquired and named 1C8H1, 2D12B6, and 3D8A10. The recombinant ScFv gene of about 750 bp was constructed. The sequencing results showed that the ScFv gene consists of a 360 bp heavy chain, a 342 bp light chain, and a 45 bp linker region. The recombinant fusion ScFv protein was expressed by IPTG induction, and a 44.5 kDa of recombinant fusion protein was obtained. In conclusion, we obtained three cell lines stably producing monoclonal antibody specifically bound to PthA-NLS, and the relative ScFv gene was constructed and successfully expressed in E. coli. These results may play an important role in further understanding the pathogenesis mechanism and in the development of possible citrus resistant to canker disease by genetic transformation and plant antibiobody.
基金supported by the Major Scientific and Technical Project in Hunan Province, China(04NK1005)
文摘Total RNA was isolated from the phloem of young shoots and retro-transcripted to cDNA by reverse transcriptasepolymerase chain reaction （RT-PCR） with specific primers; the amplified cDNA fragments were sequenced for identification of subtypes. In this study, an effective RT-PCR technique was established for detecting citrus viroids, and three citrus viroids were recognized, namely, CEVd, CVd-II, and CVd-III. The latter two were identified for the first time in China. The sequences of CVd-Ⅱ were different from all the three subtypes reported in the GenBank and seemed to be a new subtype. CVd-Ⅲb detected in the present study could be further developed as a dwarfing agent.
基金ThisprojecthadobtainedChinaPatent (No .ZL98 2 2 6 785 .1)
文摘A new technology for harnessing the dye polluted water and dye collection was developed. It is based on the enhanced evaporation by using solar, wind and air temperature energy and additional heat-electric energy. It consists of four parts: (1) evaporation carrier system (evaporation carrier and frame for evaporation carrier) for polluted water; (2)polluted water circulating system (pumping-spraying-collecting); (3)heating system; (4)workshop with polluted water reservoir-tanks and rainfall prevention roof. The polluted water was (heated in case necessary) sprayed to the evaporation carrier system and the water was evaporated when it moved in the space and downward along the carrier mainly by using natural (solar, wind and air temperature energy). In case, when there is no roof for the carrier system, the polluted water can be stored in the reservoirs (storage volume for about 20 days). The first 10-25 mm rainfall also need to be stored in the reservoirs to meet the state standard or discharging wastewater. The dye may be collected at the surface in the reservoir-tanks and the crystallized salt may be collected at the bottom plate. The black-color wastewater released by the factory is no more discharged to the surface water system of Taihu Lake Basin. About 2 kg dye and 200 kg industrial salt may be collected from each tone of the polluted water. The non-pollution production of dye may be realized by using this technology with environmental, economical and social benefits.
文摘The title compound (Ⅰ) has been synthesized by the reaction of [RuCl2(PPh3)3], closo-[B10H10]2- and PhCH(OH)COOH in (CH3)2CHOH solution. The re-action gave two main products Ⅰ and Ⅱ. Crystals of the title compound (Ⅰ) suitable fora single crystal X-ray diffraction analysis were recrystailized from n-pentane/CH2Cl2 so-lution. Its structure was determined by X-ray crystallography analysis. The crystal of{(PPh3)2RuB10 H8 [(CH3)2CHO]2} (Ⅰ) is monoclinic, space group P21/n, Mr =860. 01, with a= 12. 927(2), b= 23. 672(6), c= 14. 090(4) A, β=93. 22(2)°, V=4305 A3, Dc= 1. 327 g/cm3, Z= 4, λ(MoKα) =0. 71073A, μ=4. 6 cm-1, F(000) =1776, R = 0. 059, Rw = 0. 075, S = 2. 62. The cluster structure is based on a closoeleven-vertex {RuB10 } polyhedron skeleton with two replacing isopropoxy and twoPPh3 groups.
