AIM:Human zinc finger protein 191 (ZNF191) was cloned and characterized as a Krueppel-like transcription factor, which might be relevant to many diseases such as liver cancer,neuropsychiatric and cardiovascular diseas...AIM:Human zinc finger protein 191 (ZNF191) was cloned and characterized as a Krueppel-like transcription factor, which might be relevant to many diseases such as liver cancer,neuropsychiatric and cardiovascular diseases. Although progress has been made recently, the biological function of ZNF191 remains largely unidentified. The aim of this study was to establish a ZNF191 transgenic mouse model,which would promote the functional study of ZNF191.METHODS:Transgene fragments were microinjected into fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice.The offsprings were identified by PCR and Southern blot analysis.ZNF191 gene expression was analyzed by RT-PCR.Transgenic founder mice were used to establish transgenic mouse lineages.The first generation (F1) and the second generation (F2) mice were identified by PCR analysis.Ten-week transgenic mice were used for pathological examination.RESULTS: Four mice were identified as carrying copies of ZNF191 gene.The results of RT-PCR showed that ZNF191 gene was expressed in the liver,testis and brain in one of the transgenic mouse lineages.Genetic analysis of transgenic mice demonstrated that ZNF191 gene was integrated into the chromosome at a single site and could be transmitted stably. Pathological analysis showed that the expression of ZNF 191 did not cause obvious pathological changes in multiple tissues of transgenic mice.CONCLUSION:ZNF191 transgenic mouse model would facilitate the investigation of biological functions of ZNF191 in vivo.展开更多
AIM:To investigate the roles of c-Jun N-terminal kinase (JNK) signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS:Human gastric cancer cell lines (SGC-7901) were ...AIM:To investigate the roles of c-Jun N-terminal kinase (JNK) signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS:Human gastric cancer cell lines (SGC-7901) were treated with vitamin E succinate (VES) at 5,10,20 mg/L. Succinic acid and vitamin E were used as vehicle controls and condition medium only as an untreated (UT) control. Apoptosis was observed by 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) staining for morphological changes and by DNA fragmentation for biochemical alterations. Western blot analysis was applied to measure the expression of JNK and phosphorylated JNK.After the cells were transiently transfected with dominant negative mutant of JNK (DN- JNK) followed by treatment of VES,the expression of JNK and c-Jun protein was determined. RESULTS:The apoptotic changes were observed after VES treatment by DNA fragmentation.DNA ladder in the 20 mg/L VES group was more clearly seen than that in 10 mg/L VES group and was not detected following treatment of UT control,succinate and vitamin E.VES at 5,10 and 20 mg/L increased the expression of p-JNK by 2.5-,2.8- and 4.2- fold,respectively.VES induced the phosphorylation of JNK beginning at 1.5 h and produced a sustained increase for 24 h with the peak level at 12 h.Transient transfection of DN-JNK blocked VES-triggered apoptosis by 52%.DN-JNK significantly increased the level of JNK,while decreasing the expression of VES-induced c-Jun protein. CONCLUSION:VES-induced apoptosis in human gastric cancer SGC-7901 cells involves JNK signaling pathway via c-Jun and its downstream transcription factor.展开更多
In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a b...In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on the beads. The double strand cDNA were released from magnetic beads by digestion with NotI, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA were directionally cloned intoλExcell vector. The results show that the primary titer of the cDNA library is 7.46×106 pfu per mL and the packaging efficiency reaches 1.47×107 recombinants per 靏 DNA. λDNA extracted from two clones of plaque were digested by EcoR I and NotI, both of the clones contained inserts larger than 900 bp. These results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed.展开更多
Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, ...Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, the gene response of Populus euphratica to salt stress is discussed. It is indicated that in response to salt treatment the transcription level for some genes of Populus euphratica increases by about 1.5 times and significant difference between the responses to osmotic stress and to ion stress has been observed in gene activity.展开更多
基金Supported by the National Natural Science Foundation of China,No.39830360
文摘AIM:Human zinc finger protein 191 (ZNF191) was cloned and characterized as a Krueppel-like transcription factor, which might be relevant to many diseases such as liver cancer,neuropsychiatric and cardiovascular diseases. Although progress has been made recently, the biological function of ZNF191 remains largely unidentified. The aim of this study was to establish a ZNF191 transgenic mouse model,which would promote the functional study of ZNF191.METHODS:Transgene fragments were microinjected into fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice.The offsprings were identified by PCR and Southern blot analysis.ZNF191 gene expression was analyzed by RT-PCR.Transgenic founder mice were used to establish transgenic mouse lineages.The first generation (F1) and the second generation (F2) mice were identified by PCR analysis.Ten-week transgenic mice were used for pathological examination.RESULTS: Four mice were identified as carrying copies of ZNF191 gene.The results of RT-PCR showed that ZNF191 gene was expressed in the liver,testis and brain in one of the transgenic mouse lineages.Genetic analysis of transgenic mice demonstrated that ZNF191 gene was integrated into the chromosome at a single site and could be transmitted stably. Pathological analysis showed that the expression of ZNF 191 did not cause obvious pathological changes in multiple tissues of transgenic mice.CONCLUSION:ZNF191 transgenic mouse model would facilitate the investigation of biological functions of ZNF191 in vivo.
基金Supported by National Natural Science Foundation of China,No.39870662
文摘AIM:To investigate the roles of c-Jun N-terminal kinase (JNK) signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS:Human gastric cancer cell lines (SGC-7901) were treated with vitamin E succinate (VES) at 5,10,20 mg/L. Succinic acid and vitamin E were used as vehicle controls and condition medium only as an untreated (UT) control. Apoptosis was observed by 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) staining for morphological changes and by DNA fragmentation for biochemical alterations. Western blot analysis was applied to measure the expression of JNK and phosphorylated JNK.After the cells were transiently transfected with dominant negative mutant of JNK (DN- JNK) followed by treatment of VES,the expression of JNK and c-Jun protein was determined. RESULTS:The apoptotic changes were observed after VES treatment by DNA fragmentation.DNA ladder in the 20 mg/L VES group was more clearly seen than that in 10 mg/L VES group and was not detected following treatment of UT control,succinate and vitamin E.VES at 5,10 and 20 mg/L increased the expression of p-JNK by 2.5-,2.8- and 4.2- fold,respectively.VES induced the phosphorylation of JNK beginning at 1.5 h and produced a sustained increase for 24 h with the peak level at 12 h.Transient transfection of DN-JNK blocked VES-triggered apoptosis by 52%.DN-JNK significantly increased the level of JNK,while decreasing the expression of VES-induced c-Jun protein. CONCLUSION:VES-induced apoptosis in human gastric cancer SGC-7901 cells involves JNK signaling pathway via c-Jun and its downstream transcription factor.
基金the National Natural Science Foundation of China (Grant No. 640650)
文摘In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on the beads. The double strand cDNA were released from magnetic beads by digestion with NotI, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA were directionally cloned intoλExcell vector. The results show that the primary titer of the cDNA library is 7.46×106 pfu per mL and the packaging efficiency reaches 1.47×107 recombinants per 靏 DNA. λDNA extracted from two clones of plaque were digested by EcoR I and NotI, both of the clones contained inserts larger than 900 bp. These results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed.
基金National Natural Science Foundation of China(Grant No.39830320)
文摘Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, the gene response of Populus euphratica to salt stress is discussed. It is indicated that in response to salt treatment the transcription level for some genes of Populus euphratica increases by about 1.5 times and significant difference between the responses to osmotic stress and to ion stress has been observed in gene activity.
