Site_specific mutagenesis has been widely used in molecular biology and biochemistry. The authors have developed a simple and easy method for site_specific mutagenesis of any genes on plasmids using long distance inve...Site_specific mutagenesis has been widely used in molecular biology and biochemistry. The authors have developed a simple and easy method for site_specific mutagenesis of any genes on plasmids using long distance inverse PCR in the presence of Pfu_DNA polymerase. The efficiency of this method is higher than 90% and the entire procedure can be performed just in one tube. No subcloning is needed. This method is especially useful for obtaining mutant genes on large plasmids such as Ti plasmids used for plant transformation.展开更多
AIMS To examine the prevalance of p53 mutations in hepatocellular carcinoma (HCC) from Chongqing area and the relationship between the p53 mutations and clinicopathological features of HCC,as well as the risk factors....AIMS To examine the prevalance of p53 mutations in hepatocellular carcinoma (HCC) from Chongqing area and the relationship between the p53 mutations and clinicopathological features of HCC,as well as the risk factors. METHODS The overexpression and point mutations of tumor suppressor gene p53 in 38 cases of HCC were detected by a sensitive antigen retrieval fluid (ARF) immunohistochemical method and polymerase chain re- action(PCR)-restriction fragment length polymorphism (RFLP),and single strand conformation polymorphism (SSCP)-silver staining analysis. RESULTS The results showed that 16 of 38 HCCs had positive p53 protein (42.1%),7 HCCs had p53 mutation at 249 (18.4 % ) and 2 HCCS had point muta- tion within exon 7 other than 249. Among 9 cases of HCC with mutations,8 cases demonstrated positive p53 protein,its coincidental rate was 88.9%. The overexpression and mutations of p53 were significantly related to the differentiation and metastasis of HCCs. The frequency of p53 mutations was consistent with high prevalence of HBV and a moderate aflatoxin B1 (AFB1) exposure in our area. CONCLUSIONS The results suggest that AFB1 acts synergistically with HBV in the generation of p53 mutations. Furthermore,dietary exposure to AFB1 may mainly contribute to the tumor specific mutation at codon 249,while HBV may account for other scattered mutations in HCC.展开更多
The present article deals with thermally stratified stagnation-point flow saturated in porous medium on surface of variable thickness along with more convincing and reliable surface condition termed as melting heat tr...The present article deals with thermally stratified stagnation-point flow saturated in porous medium on surface of variable thickness along with more convincing and reliable surface condition termed as melting heat transfer.Homogeneous–heterogeneous reaction and radiative effects have been further taken into account to reconnoiterproperties of heat transfer.Melting heat transfer and phenomenon of homogeneous–heterogeneous reaction have engrossed widespread utilization in purification of metals,welding process,electroslag melting,biochemical systems,catalysis and several industrial developments.Suitable transformations are utilized to attain a scheme of ordinary differential equations possessing exceedingly nonlinear nature.Homotopic process is employed to develop convergent solutions of the resulting problem.Discussion regarding velocity,thermal field and concentration distribution for several involved parameters is pivotal part.Graphical behaviors of skin friction coefficient and Nusselt number are also portrayed.Concentration of the reactants is found to depreciate as a result of strength of both heterogeneous and homogeneous reaction parameters.With existence of melting phenomenon,declining attitude of fluid temperature is observed for higher radiation parameter.展开更多
Objective: To evaluate the incidence and pattern of rhodopsin (RHO) mutations in Chinese patients with retinitis pigmentosa (RP). Methods: Conformation sensitive gel electrophoresis (CSGE) and direct DNA sequencing we...Objective: To evaluate the incidence and pattern of rhodopsin (RHO) mutations in Chinese patients with retinitis pigmentosa (RP). Methods: Conformation sensitive gel electrophoresis (CSGE) and direct DNA sequencing were applied to detect point mutations that occurred in the five coding exons and splice sites of RHO gene in 98 index patients with RP. Results: Four patients of one ADRP family were found to have a missense mutation at codon 347, Pro347Leu. One late-onset RP patient and her daughter, without clinical expression at present, were discovered to have a novel frameshift mutation at codon 327, Pro327 (1-bp del) . Neither of the two mutations was found in 100 normal controls. Ala299Ser was found in one RP patient. Two control subjects also had Ala299Ser, suggesting its nonpathogenicity and just single nucleotide polymorphism (SNP). Conclusion: Two RP patients had rhodopsin mutations, thus the expected frequency of RHO mutations in RP is about 2.0% (95% confidence interval: 0.3%-4.4%). A highly conserved C-terminal sequence QVS(A) PA was altered due to Pro347Leu and thereby misdirecting rhodopsin to incorrect subcellular location. Loss of all phosphory-lation sites at the C-terminus and a highly conserved sequence QVS(A)PA may occur because of Pro327(1-bp del) . To elucidate the predominant biochemical defects in such mutant, transgenic mice and transfected culture cells carrying Pro327(1-bp del) would be of great value.展开更多
Abstract Four successive mass selection lines of the Pacific oyster, Crassostrea gigas, selected for faster growth in breeding pro- grams in China were examined at ten polymorphic microsatellite loci to assess the lev...Abstract Four successive mass selection lines of the Pacific oyster, Crassostrea gigas, selected for faster growth in breeding pro- grams in China were examined at ten polymorphic microsatellite loci to assess the level of allelic diversity and estimate the effective population size. These data were compared with those of their base population. The results showed that the genetic variation of the four generations were maintained at high levels with an average allelic richness of 18.8-20.6, and a mean expected heterozygosity of 0.902-0.921. They were not reduced compared with those of their base population. Estimated effective population sizes based on temporal variances in microsatellite frequencies were smaller to that of sex ratio-corrected broodstock count estimates. Using a rela- tively large number ofbroodstock and keeping an equal sex ratio in the broodstock each generation may have contributed to retaining the original genetic diversity and maintaining relatively large effective population size. The results obtained in this study showed that the genetic variation was not affected greatly by mass selection progress and high genetic variation still existed in the mass selection lines, suggesting that there is still potential for increasing the gains in future generations of C. gigas. The present study provided im- portant information for future genetic improvement by selective breeding, and for the design of suitable management guidelines for genetic breeding of C. gigas.展开更多
Based on the theory of the complex variable functions, the analysis of non-axisymmetric thermal stresses in a finite matrix containing a circular inclusion with functionally graded interphase is presented by means of ...Based on the theory of the complex variable functions, the analysis of non-axisymmetric thermal stresses in a finite matrix containing a circular inclusion with functionally graded interphase is presented by means of the least square boundary collocation technique. The distribution of thermal stress for the functionally graded interphase layer with arbitrary radial material parameters is derived by using the method of piece-wise homogeneous layers when the finite matrix is subjected to uniform heat flow. The effects of matrix size, interphase thickness and compositional gradient on the interfacial thermal stress are discussed in detail. Numerical results show that the magnitude and distribution of interfacial thermal stress in the inclusion and matrix can be designed properly by controlling these parameters.展开更多
Simultaneous effects of an on-center hydrogenic impurity and band edge non-parabolicity on intersubband optical absorption coefficients and refractive index changes of a typical GaAs/AlxGa1-xAs spherical quantum dot a...Simultaneous effects of an on-center hydrogenic impurity and band edge non-parabolicity on intersubband optical absorption coefficients and refractive index changes of a typical GaAs/AlxGa1-xAs spherical quantum dot are theoretically investigated, using the Luttinger-Kohn effective mass equation. So, electronic structure and optical properties of the system are studied by means of the matrix diagonalization technique and compact density matrix approach, respectively. Finally, effects of an impurity, band edge non-parabolicity, incident light intensity and the dot size on the linear, the third-order nonlinear and the total optical absorption coemcients and refractive index changes are investigated. Our results indicate that, the magnitudes of these optical quantities increase and their peaks shift to higher energies as the influences of the impurity and the band edge non-parabolicity are considered. Moreover, incident light intensity and the dot size have considerable effects on the optical absorption coefficients and refractive index changes.展开更多
文摘Site_specific mutagenesis has been widely used in molecular biology and biochemistry. The authors have developed a simple and easy method for site_specific mutagenesis of any genes on plasmids using long distance inverse PCR in the presence of Pfu_DNA polymerase. The efficiency of this method is higher than 90% and the entire procedure can be performed just in one tube. No subcloning is needed. This method is especially useful for obtaining mutant genes on large plasmids such as Ti plasmids used for plant transformation.
文摘AIMS To examine the prevalance of p53 mutations in hepatocellular carcinoma (HCC) from Chongqing area and the relationship between the p53 mutations and clinicopathological features of HCC,as well as the risk factors. METHODS The overexpression and point mutations of tumor suppressor gene p53 in 38 cases of HCC were detected by a sensitive antigen retrieval fluid (ARF) immunohistochemical method and polymerase chain re- action(PCR)-restriction fragment length polymorphism (RFLP),and single strand conformation polymorphism (SSCP)-silver staining analysis. RESULTS The results showed that 16 of 38 HCCs had positive p53 protein (42.1%),7 HCCs had p53 mutation at 249 (18.4 % ) and 2 HCCS had point muta- tion within exon 7 other than 249. Among 9 cases of HCC with mutations,8 cases demonstrated positive p53 protein,its coincidental rate was 88.9%. The overexpression and mutations of p53 were significantly related to the differentiation and metastasis of HCCs. The frequency of p53 mutations was consistent with high prevalence of HBV and a moderate aflatoxin B1 (AFB1) exposure in our area. CONCLUSIONS The results suggest that AFB1 acts synergistically with HBV in the generation of p53 mutations. Furthermore,dietary exposure to AFB1 may mainly contribute to the tumor specific mutation at codon 249,while HBV may account for other scattered mutations in HCC.
文摘The present article deals with thermally stratified stagnation-point flow saturated in porous medium on surface of variable thickness along with more convincing and reliable surface condition termed as melting heat transfer.Homogeneous–heterogeneous reaction and radiative effects have been further taken into account to reconnoiterproperties of heat transfer.Melting heat transfer and phenomenon of homogeneous–heterogeneous reaction have engrossed widespread utilization in purification of metals,welding process,electroslag melting,biochemical systems,catalysis and several industrial developments.Suitable transformations are utilized to attain a scheme of ordinary differential equations possessing exceedingly nonlinear nature.Homotopic process is employed to develop convergent solutions of the resulting problem.Discussion regarding velocity,thermal field and concentration distribution for several involved parameters is pivotal part.Graphical behaviors of skin friction coefficient and Nusselt number are also portrayed.Concentration of the reactants is found to depreciate as a result of strength of both heterogeneous and homogeneous reaction parameters.With existence of melting phenomenon,declining attitude of fluid temperature is observed for higher radiation parameter.
文摘Objective: To evaluate the incidence and pattern of rhodopsin (RHO) mutations in Chinese patients with retinitis pigmentosa (RP). Methods: Conformation sensitive gel electrophoresis (CSGE) and direct DNA sequencing were applied to detect point mutations that occurred in the five coding exons and splice sites of RHO gene in 98 index patients with RP. Results: Four patients of one ADRP family were found to have a missense mutation at codon 347, Pro347Leu. One late-onset RP patient and her daughter, without clinical expression at present, were discovered to have a novel frameshift mutation at codon 327, Pro327 (1-bp del) . Neither of the two mutations was found in 100 normal controls. Ala299Ser was found in one RP patient. Two control subjects also had Ala299Ser, suggesting its nonpathogenicity and just single nucleotide polymorphism (SNP). Conclusion: Two RP patients had rhodopsin mutations, thus the expected frequency of RHO mutations in RP is about 2.0% (95% confidence interval: 0.3%-4.4%). A highly conserved C-terminal sequence QVS(A) PA was altered due to Pro347Leu and thereby misdirecting rhodopsin to incorrect subcellular location. Loss of all phosphory-lation sites at the C-terminus and a highly conserved sequence QVS(A)PA may occur because of Pro327(1-bp del) . To elucidate the predominant biochemical defects in such mutant, transgenic mice and transfected culture cells carrying Pro327(1-bp del) would be of great value.
基金supported by grants from the National High Technology Research and Development Program (2012AA10A405-6)National Natural Science Foundation of China (31372524)Special Fund for Independent Innovation of Shandong Province (2013CX80202)
文摘Abstract Four successive mass selection lines of the Pacific oyster, Crassostrea gigas, selected for faster growth in breeding pro- grams in China were examined at ten polymorphic microsatellite loci to assess the level of allelic diversity and estimate the effective population size. These data were compared with those of their base population. The results showed that the genetic variation of the four generations were maintained at high levels with an average allelic richness of 18.8-20.6, and a mean expected heterozygosity of 0.902-0.921. They were not reduced compared with those of their base population. Estimated effective population sizes based on temporal variances in microsatellite frequencies were smaller to that of sex ratio-corrected broodstock count estimates. Using a rela- tively large number ofbroodstock and keeping an equal sex ratio in the broodstock each generation may have contributed to retaining the original genetic diversity and maintaining relatively large effective population size. The results obtained in this study showed that the genetic variation was not affected greatly by mass selection progress and high genetic variation still existed in the mass selection lines, suggesting that there is still potential for increasing the gains in future generations of C. gigas. The present study provided im- portant information for future genetic improvement by selective breeding, and for the design of suitable management guidelines for genetic breeding of C. gigas.
基金supported by the National Natural Science Foundation of China(Grant No.11232007)the Funding for Outstanding Doctoral Dissertation in Nanjing University of Aeronautics and Astronautics(Grant No.BCXJ11-03)Funding of Jiangsu Innovation Program for Graduate Education(Grant No.CXZZ11_0191)
文摘Based on the theory of the complex variable functions, the analysis of non-axisymmetric thermal stresses in a finite matrix containing a circular inclusion with functionally graded interphase is presented by means of the least square boundary collocation technique. The distribution of thermal stress for the functionally graded interphase layer with arbitrary radial material parameters is derived by using the method of piece-wise homogeneous layers when the finite matrix is subjected to uniform heat flow. The effects of matrix size, interphase thickness and compositional gradient on the interfacial thermal stress are discussed in detail. Numerical results show that the magnitude and distribution of interfacial thermal stress in the inclusion and matrix can be designed properly by controlling these parameters.
文摘Simultaneous effects of an on-center hydrogenic impurity and band edge non-parabolicity on intersubband optical absorption coefficients and refractive index changes of a typical GaAs/AlxGa1-xAs spherical quantum dot are theoretically investigated, using the Luttinger-Kohn effective mass equation. So, electronic structure and optical properties of the system are studied by means of the matrix diagonalization technique and compact density matrix approach, respectively. Finally, effects of an impurity, band edge non-parabolicity, incident light intensity and the dot size on the linear, the third-order nonlinear and the total optical absorption coemcients and refractive index changes are investigated. Our results indicate that, the magnitudes of these optical quantities increase and their peaks shift to higher energies as the influences of the impurity and the band edge non-parabolicity are considered. Moreover, incident light intensity and the dot size have considerable effects on the optical absorption coefficients and refractive index changes.