The location of the toxin gene of B. thuringiensis subsp. galleriae (H5ab) on the Mr-130Mdplasmid is determined by molecular cloning. Double digestion fragments (BamHⅠ and SalⅠ)and PstⅠ restriction fragments as wel...The location of the toxin gene of B. thuringiensis subsp. galleriae (H5ab) on the Mr-130Mdplasmid is determined by molecular cloning. Double digestion fragments (BamHⅠ and SalⅠ)and PstⅠ restriction fragments as well, from the 130 Md plasmid of B. thuringiensis subsp.galleriae, are ligated with the cloning vector pAT 153 respectively and transformed into E.coli strain HB 101. Out of 208 transformants, three colonies (FG2, FG9, FG19) give posi-tive hybridization reaction using the HD-1 delta-endotoxin gene as a probe. They are presum-ed to contain the delta-endotoxin gene of B. thuringiensis subsp. galleriae. Western bolt assaysindicate that Mr-130 kDal and 68 kDal, crystal proteins produced by clone FG 2 react withanticrystal protein antibody. The protein extracts of clone FG2 are lethal to Ostrinia furna-calis (Guenee). This is the first report with regard to the cloning and expression of the B. thuringiensissubsp. galleriae (H5ab) delta-endotoxin gene.展开更多
In order to reveal variation and revolution of NP genes of human avian H5 N1 influenza virus strains, the NP gene of a human avian H5 N1 influenza virus strain in Guangdong was sequenced and the global NP genes of str...In order to reveal variation and revolution of NP genes of human avian H5 N1 influenza virus strains, the NP gene of a human avian H5 N1 influenza virus strain in Guangdong was sequenced and the global NP genes of strains were retrieved. The sequences were analyzed by DNAStar 5.0, and the evolutionary speed was studied with reference to the epidemiological data. It was found that NP genes of 45 strains during 1997-2006 were homologically classified into three groups： strains in 1997-1998, strains in 2004-2005 and strains from 2003 to 2006. There were 35 substitutions in NPs in all strains accounting for a ratio of 7.03% （35/498）. An additional glycoprotein domain （NGT430-432） was found in NP genes in the strains of 2003-2006, the mutation of N370S in GD-01-06 resulted in occurrence of one more glycoprotein domain （NES368-370）. In the synonymous variation, Ks values in NP were 2.03 × 10^-5-2.55 × 10^-5 Nt/d and K. values in NP were 1.58 × 10^-6-3.10 × 10^-6 Nt/d. There didn＇t exist obviously selective pressure. An additional glycoprotein domain in every strain of 2003-2006 and one more in strain GD-01-06 might change the antigenicity of human avian H5 N1 influenza virus. The variation on human avian H5 N1 influenza strains occurred frequently in the natural world, which would result in high probability of human-human transmission along with the natural evolution of the virus.展开更多
文摘The location of the toxin gene of B. thuringiensis subsp. galleriae (H5ab) on the Mr-130Mdplasmid is determined by molecular cloning. Double digestion fragments (BamHⅠ and SalⅠ)and PstⅠ restriction fragments as well, from the 130 Md plasmid of B. thuringiensis subsp.galleriae, are ligated with the cloning vector pAT 153 respectively and transformed into E.coli strain HB 101. Out of 208 transformants, three colonies (FG2, FG9, FG19) give posi-tive hybridization reaction using the HD-1 delta-endotoxin gene as a probe. They are presum-ed to contain the delta-endotoxin gene of B. thuringiensis subsp. galleriae. Western bolt assaysindicate that Mr-130 kDal and 68 kDal, crystal proteins produced by clone FG 2 react withanticrystal protein antibody. The protein extracts of clone FG2 are lethal to Ostrinia furna-calis (Guenee). This is the first report with regard to the cloning and expression of the B. thuringiensissubsp. galleriae (H5ab) delta-endotoxin gene.
文摘In order to reveal variation and revolution of NP genes of human avian H5 N1 influenza virus strains, the NP gene of a human avian H5 N1 influenza virus strain in Guangdong was sequenced and the global NP genes of strains were retrieved. The sequences were analyzed by DNAStar 5.0, and the evolutionary speed was studied with reference to the epidemiological data. It was found that NP genes of 45 strains during 1997-2006 were homologically classified into three groups： strains in 1997-1998, strains in 2004-2005 and strains from 2003 to 2006. There were 35 substitutions in NPs in all strains accounting for a ratio of 7.03% （35/498）. An additional glycoprotein domain （NGT430-432） was found in NP genes in the strains of 2003-2006, the mutation of N370S in GD-01-06 resulted in occurrence of one more glycoprotein domain （NES368-370）. In the synonymous variation, Ks values in NP were 2.03 × 10^-5-2.55 × 10^-5 Nt/d and K. values in NP were 1.58 × 10^-6-3.10 × 10^-6 Nt/d. There didn＇t exist obviously selective pressure. An additional glycoprotein domain in every strain of 2003-2006 and one more in strain GD-01-06 might change the antigenicity of human avian H5 N1 influenza virus. The variation on human avian H5 N1 influenza strains occurred frequently in the natural world, which would result in high probability of human-human transmission along with the natural evolution of the virus.
