The biological characteristics of Lepus timibus in captivity, such as living habit, feeding habit, nutrition composition of main food, growth and development, molting rule, were studied systematically. The results showed that there were more than 30 kinds of common plants favored by L.timibus. The linear regression equation of weight gain was y=-23.223+30.929 x. The molting sequence in spring successively was head, dorsum, ear,both sides of the chest, abdomen, limbs, tails.
[Objective] The aim was to investigate the dimer formation between the movement proteins（MP）in barely yellow dwarf virus by using the technology of bimolecular fluorescence complementation technology and to further study the relationship between MP homodimerization and viral movement.[Method] The DNA sequence of bimolecular fluorescent complementary vector containing cloning multiple cloning sites,35S promoter and terminator was cloned into the expression vector pCAMBIA1300,which replicates at a higher copy number in E.coli.Then,the BYDV-MP gene fragment was amplified in the presence of the whole BYDV-PAV cDNA sequence as template and the primers designed according to the BYDV-MP gene sequence from GenBank,cloned into the modified bimolecular fluorescent complementary vectors pCAMBIA1300-NE and pCAMBIA1300-CE.The resulting vectors were transformed into Agrobacterium by electroporation method and infiltrated into the tobacco leaf.Protein interactions were observed under fluorescence microscope.[Result] Yellow fluorescence could be viewed in the leaves co-infiltrated with Agrobacterium carrying pCAMBIA1300NE-MP and pCAMBIA1300CE-MP at 2-5 d post-infiltration,while yellow fluorescence could not be observed in negative control groups.[Conclusion] BYDV-MP formed homodimers in plant cells.The results can provide theoretical basis for further in-depth research about the movement process and mechanism of BYDV.
[Objective] The paper was to study the synthesis technology of the intermediate for prothioconazole： 1-chloro-1-chloroacetyl-cyclopropane. [Method] With 2-acetylbutyrolactone as the raw material, 3,5-dichloro-2-pentanone was synthesized by chlorination and hydrolysis, and1-chloro-1-chloroacetyl-cyclopropane was synthesized by further cyclization and chlorination. [Result] The optimized synthetic process was determined by optimizing reaction conditions of each step. The overall yield of four-step reaction was 76.7%, and the product was verified by1 H NMR and13 C NMR. [Conclusion] The chlorinated product of 2-acetylbutyrolactone was directly hydrolyzed in acid solution without separation, and the final product was obtained by chlorination of cyclization product in the presence of special catalyst methanol. The whole process was smooth and the yield of the reaction was high, with high value for industrial application.
[Objective]The aim was to construct the wheat heat-shock protein 60 （HSP60） gene in prokaryotic expression vector and express HSP60 efficiently in E.coli. [Method]According to the wheat HSP60 gene sequence in GenBank,a pair of primers P1/P2 were designed and synthesized. The wheat HSP60 gene fragment was amplified from the wheat RNA by RT-PCR and inserted into bacterial expression vector of pGEX-4T-1. The construct of pGEX-4T-1-HSP60 was subsequently transformed into E.coli BL21. [Result]The construct of pGEX-4T-1-HSP60 was verified by restriction endonuclease digestion and sequenced. Compared with the sequences of wheat HSP60 genes in GenBank,homology accounted to 100%. Expression of the GST-HSP60 fusion protein was induced with IPTG. Its molecular weight was about 90 kD. The result was identified by electrophoresis of SDS-PAGE. Expression of the protein bands was consistent with the expected size. [Conclusion]The recombinant prokaryotic expression vector in pGEX-4T-1-HSP60 was constructed successfully and expressed stably in E.coli BL21. This will lay the foundation for further study on the functions of the protein and its mechanism.
The epidemiology of highly pathogenic porcine reproductive and respiratory syndrome( HP-PRRS) in Changping District,Beijing City was investigated from 2010 to 2015. Through statistics of vaccine,immune procedure and antibody levels,the epidemic prevention effect of HP-PRRS in Changping District was scientifically assessed.
