Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive pr...Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive prevention and control of PR and gradual purification of PR in different regions.展开更多
Using pseudorabies virus Ea strain as material,we inserted LacZ gene expression cassette into gE gene.After blue plaque and plaque purification,a recombinant virus PRVEa TK+-/gE+-/LacZ++ generated.Utilizing EcoR I sit...Using pseudorabies virus Ea strain as material,we inserted LacZ gene expression cassette into gE gene.After blue plaque and plaque purification,a recombinant virus PRVEa TK+-/gE+-/LacZ++ generated.Utilizing EcoR I site in LacZ gene, digested PRVEa TK+-/gE+-/LacZ++ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS,then PRVEa TK+-/gE+-/gp63+- generated after plaque purification.Four pairs of primers amplification demonstrated the virus was pure TK+-/gE+-/gp63+- mutant virus.PCR product sequence indicates there were 205bp deletion in TK gene;1247bp deletion in gE,gp63 and intergenic region of PRVEa TK+-/gE+-/gp63+- mutant virus genome DNA.Inoculation to Balb/C mice with PRVEa TK+-/gE+-/gp63+- indicates the virulence is reduced greatly.展开更多
The H2Ge=Ge：, as well as and its derivatives （X2Ge=Ge：, X=H, Me, F, C1, Br, Ph, At, ...） is a kind of new species. Its cycloaddition reactions is a new area for the study of germylene chemistry. The mechanism of t...The H2Ge=Ge：, as well as and its derivatives （X2Ge=Ge：, X=H, Me, F, C1, Br, Ph, At, ...） is a kind of new species. Its cycloaddition reactions is a new area for the study of germylene chemistry. The mechanism of the cycloaddition reaction between singlet Me2Ge=Ge： and acetaldehyde was investigated with the B3LYP/6-31G＊ method in this work. From the potential energy profile, it could be predicted that the reaction has one dominant reaction pathway. The reaction rule is that the two reactants firstly form a four-membered Ge-heterocyclic ring germylene through the [2＋2] cycloaddition reaction. Because of the 4p unoccupied orbital of Ge： atom in the four-membered Ge-heterocyclic ring germylene and the ~ orbital of acetaldehyde forming a r^--~p donor-acceptor bond, the four-membered Ge-heterocyclic ring germylene further combines with acetaldehyde to form an intermedi- ate. Because the Ge atom in intermediate happens sp3 hybridization after transition state, then, intermediate isomerizes to a spiro-Ge-heterocyclic ring compound via a transition state. The research result indicates the laws of cycloaddition reaction between Me2Ge=Ge： and ac- etaldehyde, and lays the theory foundation of the cycloaddition reaction between H2Ge=Ge： and its derivatives （X2Ge=Ge：, X=H, Me, F, C1, Br, Ph, At, ...） and asymmetric ^-bonded compounds, which are significant for the synthesis of small-ring and spiro-Ge-heterocyclic ring compounds.展开更多
基金Supported by Natural Science Foundation of Shandong Province(ZR2014CQ012)
文摘Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive prevention and control of PR and gradual purification of PR in different regions.
文摘Using pseudorabies virus Ea strain as material,we inserted LacZ gene expression cassette into gE gene.After blue plaque and plaque purification,a recombinant virus PRVEa TK+-/gE+-/LacZ++ generated.Utilizing EcoR I site in LacZ gene, digested PRVEa TK+-/gE+-/LacZ++ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS,then PRVEa TK+-/gE+-/gp63+- generated after plaque purification.Four pairs of primers amplification demonstrated the virus was pure TK+-/gE+-/gp63+- mutant virus.PCR product sequence indicates there were 205bp deletion in TK gene;1247bp deletion in gE,gp63 and intergenic region of PRVEa TK+-/gE+-/gp63+- mutant virus genome DNA.Inoculation to Balb/C mice with PRVEa TK+-/gE+-/gp63+- indicates the virulence is reduced greatly.
文摘The H2Ge=Ge：, as well as and its derivatives （X2Ge=Ge：, X=H, Me, F, C1, Br, Ph, At, ...） is a kind of new species. Its cycloaddition reactions is a new area for the study of germylene chemistry. The mechanism of the cycloaddition reaction between singlet Me2Ge=Ge： and acetaldehyde was investigated with the B3LYP/6-31G＊ method in this work. From the potential energy profile, it could be predicted that the reaction has one dominant reaction pathway. The reaction rule is that the two reactants firstly form a four-membered Ge-heterocyclic ring germylene through the [2＋2] cycloaddition reaction. Because of the 4p unoccupied orbital of Ge： atom in the four-membered Ge-heterocyclic ring germylene and the ~ orbital of acetaldehyde forming a r^--~p donor-acceptor bond, the four-membered Ge-heterocyclic ring germylene further combines with acetaldehyde to form an intermedi- ate. Because the Ge atom in intermediate happens sp3 hybridization after transition state, then, intermediate isomerizes to a spiro-Ge-heterocyclic ring compound via a transition state. The research result indicates the laws of cycloaddition reaction between Me2Ge=Ge： and ac- etaldehyde, and lays the theory foundation of the cycloaddition reaction between H2Ge=Ge： and its derivatives （X2Ge=Ge：, X=H, Me, F, C1, Br, Ph, At, ...） and asymmetric ^-bonded compounds, which are significant for the synthesis of small-ring and spiro-Ge-heterocyclic ring compounds.
