为了揭示DNA甲基化在高表达转C4-PEPC基因水稻植株对干旱耐性的作用,以高表达转玉米C4型磷酸烯醇式丙酮酸羧化酶(PEPC)基因(C4-PEPC)水稻(PC)和受体Kitaake(WT)为材料,通过发芽、水培和盆栽试验,研究了施用不同浓度的DNA甲基化抑制剂5-...为了揭示DNA甲基化在高表达转C4-PEPC基因水稻植株对干旱耐性的作用,以高表达转玉米C4型磷酸烯醇式丙酮酸羧化酶(PEPC)基因(C4-PEPC)水稻(PC)和受体Kitaake(WT)为材料,通过发芽、水培和盆栽试验,研究了施用不同浓度的DNA甲基化抑制剂5-氮杂胞苷(5-azaC)联合干旱处理下,水稻芽期、苗期和生育后期,种子发芽率,苗期叶片的相对含水量、丙二醛、脯氨酸、总可溶性糖及其组分及总可溶性蛋白含量、PEPC酶活性以及C4-PEPC、与蔗糖非发酵1(SNF1)相关蛋白激酶(SNF1-related protein kinase 1s,SnRK1s)基因以及甲基转移酶基因表达的变化、剑叶的光合参数,并最后考察产量构成因子的变化,结果表明:5-azaC对水稻芽期和苗期干旱胁迫作用浓度呈现剂量效应,即低浓度促进,而高浓度抑制,其中作用浓度:芽期<苗期;而对芽期和苗期有促进和抑制的2个浓度外施于水稻生育后期的水稻植株后,则出现恶化的效应,表现为矮化、剑叶的净光合速率和单株产量下降。在缓解干旱抑制的浓度下,PC苗期的缓解效果好于WT,这种差异的表现与其内源蔗糖、SnRK1s基因以及OsMET1b的表达差异同步,而且PC叶片C4-PEPC表达对不同浓度的5-azaC处理也呈现剂量效应。综上,DNA甲基化参与了水稻干旱响应,但不同生育期表现不同,其中糖信号在调节DNA甲基化增强PC干旱耐性起重要作用。展开更多
Objective: To select the ovarian carcinoma cell lines with high frequent metastasis and study the association between nm23-H1 gene expression and metastasis of ovarian carcinoma. Methods: Each ovarian cancer cell line...Objective: To select the ovarian carcinoma cell lines with high frequent metastasis and study the association between nm23-H1 gene expression and metastasis of ovarian carcinoma. Methods: Each ovarian cancer cell line was transplanted subcutaneously into the flank of nude mice, and the metastatic behavior was evaluated by counting lung tumor foci at different time points. The metastatic tumors were cultured in vitro, then substrain was established and transplanted subcutaneously three times. The RNA level of nm23 in 8 human ovarian cancer cell lines were examined by northern-blot. Results: Of the 8 human ovarian cancer cell lines, 4 had high requent metastatic potentiality. The expression of nm23 RNA in human ovarian cancer cells was inversely related to metastatic behavior in the experimental animals (r=0.96, P=0.0001). Conclusion: The difference of the tendency of metastasis which was determined by genetic and molecular levels was significant among different type of cell lines and subtypes. The expression of nm23 mRNA in human ovarian carcinomas was correlated closely with the reduced metastatic behavior in experimental animals and may serve as a sensitive prognostic indicator for ovarian cancer.展开更多
目的研究染矽尘大鼠毒理效应相关差异基因表达谱。方法应用气管暴露法对雄性Wistar大鼠进行二氧化硅（50mg／ml）染尘，对照组大鼠气管内灌注1ml灭菌生理盐水。染尘后第14天处死大鼠，取肺组织。应用Ⅲumina激光共聚焦光纤微珠基因芯片...目的研究染矽尘大鼠毒理效应相关差异基因表达谱。方法应用气管暴露法对雄性Wistar大鼠进行二氧化硅（50mg／ml）染尘，对照组大鼠气管内灌注1ml灭菌生理盐水。染尘后第14天处死大鼠，取肺组织。应用Ⅲumina激光共聚焦光纤微珠基因芯片技术，研究大鼠肺组织差异基因表达谱，采用因注释、富集数据库（thedatabase for annotation，visualization and integrated discovery，DAVID）生物信息学分析工具，对差异基因的基因功能和生物学通路进行富集统计学分析。结果检测出的有效基因共22107个，筛选出染矽尘组与对照组的差异基因共1567个，其中上调基因为765个，下调基因为802个。与毒理效应相关的差异基因有461个，其中上调基因为285个，下调的有176个。Real—timePCR实验验证了血红素加氧酶（HMOX1）、过氧化物歧化酶（SOD1）基因上调的趋势和基因芯片结果是一致的。结论二氧化硅粉尘诱导的大鼠肺组织纤维化中起毒理效应的大量相关基因上调或下调，二氧化硅所致的肺纤维化可能存在复杂的基因网络调控系统，其中，毒理机制是肺纤维化发生发展过程中的重萼一环。展开更多
[Objective] The study was to investigate roles of Brassica napus EINB in （ BnEIN3 ） resistance to Sclerotinia sclerotiorum. [ Methods] Genomic PCR and RT-PCR were carded out to isolate genomic DNA and cDNA sequences...[Objective] The study was to investigate roles of Brassica napus EINB in （ BnEIN3 ） resistance to Sclerotinia sclerotiorum. [ Methods] Genomic PCR and RT-PCR were carded out to isolate genomic DNA and cDNA sequences of BnEIN3 from oilseed rape, based on the highly conserved region of EIN3 gene from Arabidopsis thaliana and the homologous sequences of oilseed rape ESTs. Expression levels of BnEIN3 were detected in three varieties of oilseed rape inoculated with S. sclerotiorum by real-time quantitative PCR.[Results] A 1 947 bp DNA fragment was obtained from oilseed rape. The fragment shared 82% identity to A. thaliana EIIV3, encoded 614 amino acids containing an EIN3 domain, and was named as BnEIN3. Real-time PCR results showed that expression patterns of BnEIN3 were drastically different in different varieties. In highly resistant oilseed rape variety D083, BnEIN3 expression level was significantly increased 72 h after S. sclerotiorum inoculation whereas in middle resistant and susceptible varieties Zhongshuang 9 and 84039, BnEIN3 expression was suppressed. [ Conclusion ] BnEIIV3 may play an important role in oilseed rape resistance to S. sclerotiorum.展开更多
Maize （Zea raays L.） is one of the most important crops because of the remarkable properties of its hybrid, which is responsible for the high commercial value of hybrid maize. The genetic basis of heterosis （hybrid...Maize （Zea raays L.） is one of the most important crops because of the remarkable properties of its hybrid, which is responsible for the high commercial value of hybrid maize. The genetic basis of heterosis （hybrid vigor） is not well understood. A differential display technique was performed to identify genes with differential expression across twelve maize inbred lines and thirty-three hybrids during ear development. An incomplete diallel design was used to investigate the relationship between the global framework of differential gene expression and heterosis. It was found that the genes belonging to MONO pattern （i.e., genes expressed in both parental lines and in hybrid） was the highest in percentage among the total five patterns and illustrated that the properties of differentially expressed genes are not entirely responsible for heterosis. Furthermore,a larger number of differentially expressed genes in hybrid, which serves as a major reservoir for generating novel phenotypes that exhibit heterosis of certain agronomic traits during early development and differentiation of maize ear. Moreover, there were some silent genesin hybrids that are responsible for the arrest or abortion of spikelets and for the increase in kernels weight.展开更多
Objectives: In order to investigate the role of mono-cytes and human leukocyte antigen (HLA) class II an-tigen in herpes simplex virus-2 (HSV-2) infection, westudied the effect of HSV-2 infection in vitro on theexpres...Objectives: In order to investigate the role of mono-cytes and human leukocyte antigen (HLA) class II an-tigen in herpes simplex virus-2 (HSV-2) infection, westudied the effect of HSV-2 infection in vitro on theexpression of HLA class II antigen on monocytes.Methods: Monocytes were infected with HSV-2(Strain 333). Culture cells were collected 1, 3, 5 and 7days after infection. The levels of expression of HLAclass II antigen were measured by using alkaline phos-phatase antialkaline phosphatase method (APAAP).Results: The levels of the expression of HLA class IIantigen on monocytes significantly decreased on thefirst day of post-infection, and then gradually returnedto levels found in the controls by the 7th day post-infection.Conclusion: HLA class II antigen expression onmonocytes was inhibited with HSV-2 infection, whichmight be one means of virus escape at an early phase.The expression of HLA class II antigen may play animportant role in herpes simplex viurs-2 pathogenic-ity and immunity.展开更多
文摘为了揭示DNA甲基化在高表达转C4-PEPC基因水稻植株对干旱耐性的作用,以高表达转玉米C4型磷酸烯醇式丙酮酸羧化酶(PEPC)基因(C4-PEPC)水稻(PC)和受体Kitaake(WT)为材料,通过发芽、水培和盆栽试验,研究了施用不同浓度的DNA甲基化抑制剂5-氮杂胞苷(5-azaC)联合干旱处理下,水稻芽期、苗期和生育后期,种子发芽率,苗期叶片的相对含水量、丙二醛、脯氨酸、总可溶性糖及其组分及总可溶性蛋白含量、PEPC酶活性以及C4-PEPC、与蔗糖非发酵1(SNF1)相关蛋白激酶(SNF1-related protein kinase 1s,SnRK1s)基因以及甲基转移酶基因表达的变化、剑叶的光合参数,并最后考察产量构成因子的变化,结果表明:5-azaC对水稻芽期和苗期干旱胁迫作用浓度呈现剂量效应,即低浓度促进,而高浓度抑制,其中作用浓度:芽期<苗期;而对芽期和苗期有促进和抑制的2个浓度外施于水稻生育后期的水稻植株后,则出现恶化的效应,表现为矮化、剑叶的净光合速率和单株产量下降。在缓解干旱抑制的浓度下,PC苗期的缓解效果好于WT,这种差异的表现与其内源蔗糖、SnRK1s基因以及OsMET1b的表达差异同步,而且PC叶片C4-PEPC表达对不同浓度的5-azaC处理也呈现剂量效应。综上,DNA甲基化参与了水稻干旱响应,但不同生育期表现不同,其中糖信号在调节DNA甲基化增强PC干旱耐性起重要作用。
基金This work was supported by the grants from 973 National Great Foundation Research Program of China(No.2002CB513100)the National Prominent Youth Foundation of China(No.30025017).
文摘Objective: To select the ovarian carcinoma cell lines with high frequent metastasis and study the association between nm23-H1 gene expression and metastasis of ovarian carcinoma. Methods: Each ovarian cancer cell line was transplanted subcutaneously into the flank of nude mice, and the metastatic behavior was evaluated by counting lung tumor foci at different time points. The metastatic tumors were cultured in vitro, then substrain was established and transplanted subcutaneously three times. The RNA level of nm23 in 8 human ovarian cancer cell lines were examined by northern-blot. Results: Of the 8 human ovarian cancer cell lines, 4 had high requent metastatic potentiality. The expression of nm23 RNA in human ovarian cancer cells was inversely related to metastatic behavior in the experimental animals (r=0.96, P=0.0001). Conclusion: The difference of the tendency of metastasis which was determined by genetic and molecular levels was significant among different type of cell lines and subtypes. The expression of nm23 mRNA in human ovarian carcinomas was correlated closely with the reduced metastatic behavior in experimental animals and may serve as a sensitive prognostic indicator for ovarian cancer.
文摘目的研究染矽尘大鼠毒理效应相关差异基因表达谱。方法应用气管暴露法对雄性Wistar大鼠进行二氧化硅（50mg／ml）染尘，对照组大鼠气管内灌注1ml灭菌生理盐水。染尘后第14天处死大鼠，取肺组织。应用Ⅲumina激光共聚焦光纤微珠基因芯片技术，研究大鼠肺组织差异基因表达谱，采用因注释、富集数据库（thedatabase for annotation，visualization and integrated discovery，DAVID）生物信息学分析工具，对差异基因的基因功能和生物学通路进行富集统计学分析。结果检测出的有效基因共22107个，筛选出染矽尘组与对照组的差异基因共1567个，其中上调基因为765个，下调基因为802个。与毒理效应相关的差异基因有461个，其中上调基因为285个，下调的有176个。Real—timePCR实验验证了血红素加氧酶（HMOX1）、过氧化物歧化酶（SOD1）基因上调的趋势和基因芯片结果是一致的。结论二氧化硅粉尘诱导的大鼠肺组织纤维化中起毒理效应的大量相关基因上调或下调，二氧化硅所致的肺纤维化可能存在复杂的基因网络调控系统，其中，毒理机制是肺纤维化发生发展过程中的重萼一环。
文摘[Objective] The study was to investigate roles of Brassica napus EINB in （ BnEIN3 ） resistance to Sclerotinia sclerotiorum. [ Methods] Genomic PCR and RT-PCR were carded out to isolate genomic DNA and cDNA sequences of BnEIN3 from oilseed rape, based on the highly conserved region of EIN3 gene from Arabidopsis thaliana and the homologous sequences of oilseed rape ESTs. Expression levels of BnEIN3 were detected in three varieties of oilseed rape inoculated with S. sclerotiorum by real-time quantitative PCR.[Results] A 1 947 bp DNA fragment was obtained from oilseed rape. The fragment shared 82% identity to A. thaliana EIIV3, encoded 614 amino acids containing an EIN3 domain, and was named as BnEIN3. Real-time PCR results showed that expression patterns of BnEIN3 were drastically different in different varieties. In highly resistant oilseed rape variety D083, BnEIN3 expression level was significantly increased 72 h after S. sclerotiorum inoculation whereas in middle resistant and susceptible varieties Zhongshuang 9 and 84039, BnEIN3 expression was suppressed. [ Conclusion ] BnEIIV3 may play an important role in oilseed rape resistance to S. sclerotiorum.
文摘Maize （Zea raays L.） is one of the most important crops because of the remarkable properties of its hybrid, which is responsible for the high commercial value of hybrid maize. The genetic basis of heterosis （hybrid vigor） is not well understood. A differential display technique was performed to identify genes with differential expression across twelve maize inbred lines and thirty-three hybrids during ear development. An incomplete diallel design was used to investigate the relationship between the global framework of differential gene expression and heterosis. It was found that the genes belonging to MONO pattern （i.e., genes expressed in both parental lines and in hybrid） was the highest in percentage among the total five patterns and illustrated that the properties of differentially expressed genes are not entirely responsible for heterosis. Furthermore,a larger number of differentially expressed genes in hybrid, which serves as a major reservoir for generating novel phenotypes that exhibit heterosis of certain agronomic traits during early development and differentiation of maize ear. Moreover, there were some silent genesin hybrids that are responsible for the arrest or abortion of spikelets and for the increase in kernels weight.
文摘Objectives: In order to investigate the role of mono-cytes and human leukocyte antigen (HLA) class II an-tigen in herpes simplex virus-2 (HSV-2) infection, westudied the effect of HSV-2 infection in vitro on theexpression of HLA class II antigen on monocytes.Methods: Monocytes were infected with HSV-2(Strain 333). Culture cells were collected 1, 3, 5 and 7days after infection. The levels of expression of HLAclass II antigen were measured by using alkaline phos-phatase antialkaline phosphatase method (APAAP).Results: The levels of the expression of HLA class IIantigen on monocytes significantly decreased on thefirst day of post-infection, and then gradually returnedto levels found in the controls by the 7th day post-infection.Conclusion: HLA class II antigen expression onmonocytes was inhibited with HSV-2 infection, whichmight be one means of virus escape at an early phase.The expression of HLA class II antigen may play animportant role in herpes simplex viurs-2 pathogenic-ity and immunity.
