为利用河套蜜瓜为生物反应器生产可用于治疗肝病的蛋白因子,以人胎肝mRNA为模板,经RT-PCR扩增获得了471 bp cDNA片段。克隆到pMD18-T载体,命名为pMD-ALR,测序分析结果与GenBank中报道的hALR编码序列完全相同。酶切回收hALR片段,插入到...为利用河套蜜瓜为生物反应器生产可用于治疗肝病的蛋白因子,以人胎肝mRNA为模板,经RT-PCR扩增获得了471 bp cDNA片段。克隆到pMD18-T载体,命名为pMD-ALR,测序分析结果与GenBank中报道的hALR编码序列完全相同。酶切回收hALR片段,插入到含有甜瓜果实黄瓜素(Cucumisin)基因启动子的果实特异性表达载体pPZP-CGN中,构建了hALR甜瓜果实特异性表达载体pPZP-CAN。花粉管通道法转化试验大田自花授粉花63朵,收获T0代果实27颗,结实率为42.9%。提取T0代种子无菌苗DNA,经PCR、PCR-southern和斑点杂交检测有13株呈阳性。展开更多
Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripoten...Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripotency.However,the presence and function of pluripotency-associated lincRNAs in cancer cells with pluripotency features are unknown.In this study,we used embryonal carcinoma(EC)P19 cell lines to investigate the expression level of Halr1 in pluripotency and retinoic acid(RA)-induced differentiated states.Down-regulation of pluripotency associated factors such as OCT4,NANOG,SSEA1 and alkaline phosphatase at transcript and protein levels were used to confirm the differentiated status of P19 cells.Quantitative measurement of Halr1 transcript levels revealed a 79% decrease during RA-induced differentiation of P19 cells.These results indicate that upon exiting the pluripotency state the expression level of Halr1 similar to core pluripotency factors is remarkably reduced.展开更多
利用三亲融合法将含有人肝再生增强因子(hum an augm en ter of liver regeneration,hALR)的果实特异性表达载体pPZP-CAN导入农杆菌LBA 4404,转化河套蜜瓜子叶外植体,经诱导与筛选获得了抗性再生植株,PCR扩增、PCR-Sou thern和斑点杂交...利用三亲融合法将含有人肝再生增强因子(hum an augm en ter of liver regeneration,hALR)的果实特异性表达载体pPZP-CAN导入农杆菌LBA 4404,转化河套蜜瓜子叶外植体,经诱导与筛选获得了抗性再生植株,PCR扩增、PCR-Sou thern和斑点杂交检测证明hALR已整合入河套蜜瓜再生植株基因组中.展开更多
文摘为利用河套蜜瓜为生物反应器生产可用于治疗肝病的蛋白因子,以人胎肝mRNA为模板,经RT-PCR扩增获得了471 bp cDNA片段。克隆到pMD18-T载体,命名为pMD-ALR,测序分析结果与GenBank中报道的hALR编码序列完全相同。酶切回收hALR片段,插入到含有甜瓜果实黄瓜素(Cucumisin)基因启动子的果实特异性表达载体pPZP-CGN中,构建了hALR甜瓜果实特异性表达载体pPZP-CAN。花粉管通道法转化试验大田自花授粉花63朵,收获T0代果实27颗,结实率为42.9%。提取T0代种子无菌苗DNA,经PCR、PCR-southern和斑点杂交检测有13株呈阳性。
基金supported by a Ferdowsi University of Mashhad grant to HD(No.22534).
文摘Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripotency.However,the presence and function of pluripotency-associated lincRNAs in cancer cells with pluripotency features are unknown.In this study,we used embryonal carcinoma(EC)P19 cell lines to investigate the expression level of Halr1 in pluripotency and retinoic acid(RA)-induced differentiated states.Down-regulation of pluripotency associated factors such as OCT4,NANOG,SSEA1 and alkaline phosphatase at transcript and protein levels were used to confirm the differentiated status of P19 cells.Quantitative measurement of Halr1 transcript levels revealed a 79% decrease during RA-induced differentiation of P19 cells.These results indicate that upon exiting the pluripotency state the expression level of Halr1 similar to core pluripotency factors is remarkably reduced.
文摘利用三亲融合法将含有人肝再生增强因子(hum an augm en ter of liver regeneration,hALR)的果实特异性表达载体pPZP-CAN导入农杆菌LBA 4404,转化河套蜜瓜子叶外植体,经诱导与筛选获得了抗性再生植株,PCR扩增、PCR-Sou thern和斑点杂交检测证明hALR已整合入河套蜜瓜再生植株基因组中.
