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121.
HAN Ya-ling ZHAO Xin YAN Cheng-hui KANG Jian ZHANG Xiao-lin DENG Jie XU Hong-mei LIU Hai-wei 《园艺学报》2008,24(8):1483-1489
AIM: In order to explore the regulatory mechanisms of the human cellular repressor of E1A-stimulated genes (hCREG1) in vascular smooth muscle cells (VSMCs)-HITASY and in human umbilical vein endothelial cells (HUVECs), we clone and construct hCREG1 promoter vector to detect its expression in different vascular cells. METHODS: With the results of biological information, the fragments including -3 677 bp, -2 310 bp and -945 bp upstream sequences of hCREG1 were amplified respectively using human genomic DNA template by PCR. The products were inserted into pMD18-T vector, and then were subcloned into pEGFP-1 report vector to obtain the pEGFP-hCREG1-promoter vectors. The pEGFP-hCREG1-P3677, pEGFP-hCREG1-P2310 and pEGFP-hCREG1-P945 vectors were transfected into HITASY cells and HUVECs transiently and the expression of green fluorescent protein (GFP) was detected respectively by Western blotting and fluorescent microscope. RESULTS: It was confirmed that all three PCR fragments inserted into vectors were corrected by sequencing analysis. However, the expression of GFP was different significantly in both types of vascular cells. The expression of GFP in HUVECs was higher than that in HITASY cells. Meanwhile, the expression of GFP in HITASY cells with 0.5% FBS was increased obviously compared to that in HITASY cells with 10% FBS. CONCLUSION: The reporter vectors of hCREG1 promoter are constructed successfully in which the core promoter region might be located in -945 bp-0 bp of 5′ upstream sequences. This study will provide an experimental basis for exploring the regulation of hCREG1 expression. 相似文献
122.
AIM: To study the role of gap junction(GJ) in the pathogenesis of epilepsy. METHODS: The expression of connexin(Cx) 32 and Cx43 was evaluated in penlylenetetrazol(PTZ)-induced epilepsy rats at different time points with immunohistochemistry and real-time RT-PCR. The uncoupler of gap junction,carbenoxolone (CBX), and anti-epilepsy drug,carbamazepine (CBZ), were introduced to investigate the role of gap junction in epilepsy. RESULTS: The expression of Cx32 in the cortex and hippocampus of rats increased 2 h after PTZ administration, and was more obvious 8 h later, so was the expression of Cx43. CBX not only notably inhibited the expression of Cx32 and Cx43, but also suppressed the seizures. CBZ had no obvious effect on Cx32/43 expression. Real-time RT-PCR examination showed that the level of Cx32 mRNA went up rapidly 2 h after seizures, and began to decrease 10 h later. The level of Cx43 mRNA in PTZ group was higher than that in control group from 2 h to 5 h. CBX inhibited both the seizures and the increase in the mRNA expression of Cx32/43, while CBZ only suppressed the seizures and did not affect the expression of Cx32/43. CONCLUSION: GJ between neurons is reinforced after epileptic activities and takes part in the pathophysiological mechanism of synchronization and epilepsy. CBZ has no effect on the expression of Cx32 and Cx43, indicating that the anti-epileptic mechanism of CBZ is independent of GJ. 相似文献
123.
黄瓜叶面积测量方法的评价 总被引:4,自引:0,他引:4
采用方格纸法、等腰三角形法及数码相机拍照法测定黄瓜叶面积。结果表明,采用这3种方法测定的结果差异不显著。综合考虑耗时、信息量、先进性等因素,数码相机拍照法为最优测定方法。 相似文献
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以紫花苜蓿"大叶苜蓿""维多利亚""阿尔冈金"为试材,采用土培法,研究了硒胁迫对3种紫花苜蓿叶片酶促防御系统的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)活性及丙二醛(MDA)、可溶性蛋白质含量的影响。结果表明:在100μmol·L-1Se时,硒胁迫显著提高了3种紫花苜蓿叶片SOD、APX、POD、CAT活性和可溶性蛋白质含量。Se为900μmol·L-1时,SOD、APX活性和可溶性蛋白质、MDA含量显著增加,而CAT、POD活性显著降低。不同品种间抗氧化酶活性相比,"大叶苜蓿"抗氧化酶SOD、APX、POD、CAT活性显著高于其它2个苜蓿品种,且差异显著。综合评价表明,"大叶苜蓿"的抗氧化能力最强,其次为"阿尔冈金","维多利亚"苜蓿的抗氧化能力最差。 相似文献
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以亲缘关系极近的新高系梨10个品种及被视为‘新高’亲本的‘天之川’和‘今村秋’梨等为试验材料,对其基因组DNA进行了PCR-RFLP检测、DNA序列测定及生物信息学分析。结果表明:新高系梨10个品种即‘新高’、‘黄金’、‘水晶’、‘早生黄金’、‘早蜜新高’、‘天皇’、‘金秋’、‘晚大新高’、‘农家新高’和‘鲜黄’的S基因型分别为S3S9、S3S4、S3S9、S3S4、S3S9、S3S9、S3S9、S3S9、S3S9和S3S5;‘天之川’、‘今村秋’的S基因型分别为S1S9和S1S6,通过基因型判定‘今村秋’非‘新高’梨的父本。 相似文献
130.