目的 观察损伤对成年动物骨骼肌卫星细胞增殖的影响 ,以获取基因治疗自体移植的载体。 方法 以阳离子脂质体介导法将 p RSV- L ac Z或 p Rch- TH质粒转入培养肌细胞。 结果 骨骼肌损伤后 ,分离出的肌卫星细胞及组织块游离成肌细胞...目的 观察损伤对成年动物骨骼肌卫星细胞增殖的影响 ,以获取基因治疗自体移植的载体。 方法 以阳离子脂质体介导法将 p RSV- L ac Z或 p Rch- TH质粒转入培养肌细胞。 结果 骨骼肌损伤后 ,分离出的肌卫星细胞及组织块游离成肌细胞数均显著增高 ;外源基因能在细胞内表达。 结论 损伤可诱导成年骨骼肌卫星细胞增殖并促进体外培养肌细胞成活。展开更多
The nudeotide sequence of the nifLA promoter of Enterobacter cloacae E26 was determined, and the transcription start site of nifLA was mapped by the primer extension. Studies on the oxygen regulation of E. cloacae nif...The nudeotide sequence of the nifLA promoter of Enterobacter cloacae E26 was determined, and the transcription start site of nifLA was mapped by the primer extension. Studies on the oxygen regulation of E. cloacae nifLA promoter with the nifL’-lacZ fusion showed that the nifLA promoter was sensitive to oxygen as Kebsiella pneumoniae nifLA promoter reported previously. Comparison between the nifLA promoter sequences of E. cloacae and K. pneumoniae revealed the presence of an 11-bp conserved sequence between the - 24/ -12 consensus of σ54-dependent promoter and NtrC binding motif. The 11-bp sequence is speculated to be involved in the oxygen regulation of the nifLA promoter of both enteric bacteria.展开更多
To investigate whether a reporter LacZ gene could be transferred into cultured ocular cells of human eyes in vitro Methods Fibroblast cells of Tenon's capsule, trabecular meshwork cells, and muscle celms in ...To investigate whether a reporter LacZ gene could be transferred into cultured ocular cells of human eyes in vitro Methods Fibroblast cells of Tenon's capsule, trabecular meshwork cells, and muscle celms in the ciliary body of human eyes were cultured and the pcDNA3 LacZ gene was transferred into these cells using a cationic liposome delivery system The cells were subsequently fixed with 4% paraformaldehyde, mixed with X gal, then observed under a microscope Results Blue stain was seen in the cytoplasm of the cultured cells under the microscope, demonstrating the successful transfer of the LacZ gene into these cells Conclusion Reporter LacZ gene was easily transferred into the cultured ocular cells in vitro This provides insights into the transfer of the genes into these cells to study the pathogenesis and therapy of glaucoma展开更多
Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells...Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells incorporated with exogenous gene(lacZ constructs encoding Escherichia coliβ-galactosidase:β-gal)were intro-duced into chick embryos by the injection of cells into stage X blastoderm.Manipulated eggs were incubated for 3(trial1)or 6(trial 2)days,and the expression of lacZ DNA was detected by a histochemical staining method ofβ-galactosi-dase and polymerase chain reaction(PCR)analysis.Results:The survival rates of the manipulated embryos incu-bated for 3 days(stage 18-20:trial 1)and 6 days(stage 28,30:trial 2)were about 42%and 38%,respectively.The expression rates of the lacZ gene in the embryos in the trials 1 and 2 were about 60%and 23%,respectively,forthe survived embryos.Conclusion:The rate of embryonic viability and expression rate of introduced genes were notso high,but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chickembryos.展开更多
文摘目的 观察损伤对成年动物骨骼肌卫星细胞增殖的影响 ,以获取基因治疗自体移植的载体。 方法 以阳离子脂质体介导法将 p RSV- L ac Z或 p Rch- TH质粒转入培养肌细胞。 结果 骨骼肌损伤后 ,分离出的肌卫星细胞及组织块游离成肌细胞数均显著增高 ;外源基因能在细胞内表达。 结论 损伤可诱导成年骨骼肌卫星细胞增殖并促进体外培养肌细胞成活。
基金Project supported by the National Natural Science Foundation of China.
文摘The nudeotide sequence of the nifLA promoter of Enterobacter cloacae E26 was determined, and the transcription start site of nifLA was mapped by the primer extension. Studies on the oxygen regulation of E. cloacae nifLA promoter with the nifL’-lacZ fusion showed that the nifLA promoter was sensitive to oxygen as Kebsiella pneumoniae nifLA promoter reported previously. Comparison between the nifLA promoter sequences of E. cloacae and K. pneumoniae revealed the presence of an 11-bp conserved sequence between the - 24/ -12 consensus of σ54-dependent promoter and NtrC binding motif. The 11-bp sequence is speculated to be involved in the oxygen regulation of the nifLA promoter of both enteric bacteria.
文摘To investigate whether a reporter LacZ gene could be transferred into cultured ocular cells of human eyes in vitro Methods Fibroblast cells of Tenon's capsule, trabecular meshwork cells, and muscle celms in the ciliary body of human eyes were cultured and the pcDNA3 LacZ gene was transferred into these cells using a cationic liposome delivery system The cells were subsequently fixed with 4% paraformaldehyde, mixed with X gal, then observed under a microscope Results Blue stain was seen in the cytoplasm of the cultured cells under the microscope, demonstrating the successful transfer of the LacZ gene into these cells Conclusion Reporter LacZ gene was easily transferred into the cultured ocular cells in vitro This provides insights into the transfer of the genes into these cells to study the pathogenesis and therapy of glaucoma
基金financially supported by the Ministry of Education,Science and Culture,Japanthe Society for the Promotion of Science(JSPS)+1 种基金the Sumitomo Foundationthe Nissan Science Foundation
文摘Aim:To examine the transfection of exogenous genes into chick embryos,applying the characteristics of avianleukosis vires(ALV)-induced chicken B cell line DT40 to the production of chimeric birds.Methods:The DT40cells incorporated with exogenous gene(lacZ constructs encoding Escherichia coliβ-galactosidase:β-gal)were intro-duced into chick embryos by the injection of cells into stage X blastoderm.Manipulated eggs were incubated for 3(trial1)or 6(trial 2)days,and the expression of lacZ DNA was detected by a histochemical staining method ofβ-galactosi-dase and polymerase chain reaction(PCR)analysis.Results:The survival rates of the manipulated embryos incu-bated for 3 days(stage 18-20:trial 1)and 6 days(stage 28,30:trial 2)were about 42%and 38%,respectively.The expression rates of the lacZ gene in the embryos in the trials 1 and 2 were about 60%and 23%,respectively,forthe survived embryos.Conclusion:The rate of embryonic viability and expression rate of introduced genes were notso high,but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chickembryos.
