Cu2＋胁迫对凤眼莲根系活力及Cu2＋-ATPase 活性的影响

通过水培法研究了不同Cu2＋浓度（0．5,1．0,2．5,5．0,10．0 mg/L）对水生植物凤眼莲[Eichhornia crassipes]蛋白质含量、根系活力及Cu2＋-ATP酶活性的影响．结果表明,Cu2＋胁迫对凤眼莲蛋白质含量、根系活力及Cu2＋-ATP酶活性的影响与 Cu2＋胁迫浓度及胁迫时间均有关,在 Cu2＋胁迫时间为1,3,10 d 时,各Cu2＋胁迫浓度下均有利于凤眼莲叶片蛋白质含量的积累．一定浓度和时间的 Cu2＋胁迫使凤眼莲体内蛋白质含量应激积累增多．在Cu2＋胁迫时间为1 d时,随着Cu2＋浓度的增加,凤眼莲根系活力逐渐增大;当胁迫时间达10 d时,根系活力在Cu2＋浓度为1．0 mg/L Cu2＋时达最大,以后逐渐降低．试验表明,随着 Cu2＋浓度的增加,叶片Cu2＋-ATP酶活性在0．5 mg/L时达最大,之后逐渐降低;随Cu2＋浓度的增加根系Cu2＋-ATP酶活性的变化趋势与叶片Cu2＋-ATP酶活性的变化趋势相似．     

少棘蜈蚣候选毒素多肽κ-SLPTX-Ssm1b的原核表达与纯化

κ-SLPTX-Ssm1b是从少棘蜈蚣的c DNA文库中获得的一类功能未知的毒素,它与钾通道抑制剂κ-SLPTX-Ssm1a高度同源,由50个氨基酸残基构成,含有3对二硫键的多肽分子.利用p ET-43-His-SUMO原核表达载体,通过E.coli SHuffleTM菌种自动诱导表达,随后利用镍柱和RP-HPLC纯化,并通过MALDI-TOF质谱鉴定其分子量为5532.1870 Da,与理论值5532.3 Da相符.其产量为1 mg/L,为进一步鉴定其电生理活性奠定了基础.     

昆虫神经毒素AaIT原核表达、纯化及抗体制备

人工合成Aa IT的成熟基因,连接到p ET32载体并转化大肠杆菌,IPTG诱导表达,纯化重组Aa IT蛋白并免疫小鼠,经蛋白质-G柱对抗血清进行纯化.研究结果表明,纯化得到了Aa IT蛋白经免疫小鼠和纯化抗血清得到了特异性强的Aa IT抗体蛋白.     

人超氧化物歧化酶（SOD）基因的原核表达与蛋白纯化

从人直肠癌细胞株Colo320中提取总RNA,经RT-PCR扩增出SOD基因片段,经限制性内切酶（BamHI,Sinai）酶切后按正确的读码框顺序插入到pGEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCR和质粒双酶切鉴定、序列测定确认,证实成功地构建了人SOD基因融合表达载体．转化菌经IPTG诱导表达,SDS-PAGE显示有与预期大小约44kD相吻合的融合蛋白带,获得了可溶性表达的目的蛋白,采用Glutathione Sepha—rose4B亲和层析对重组蛋白进行纯化,获得了纯度很高的目的蛋白．     

骨形态发生蛋白-2在大肠杆菌中的表达与纯化

骨形态发生蛋白-2（bone morphogenetic protein,BMP-2）具有多种生物活性,有潜在的药用价值.为探索利用基因工程技术重组BMP-2并在大肠杆菌表达系统表达的可行性.在大肠杆菌中重组和筛选BMp-2基因,SDS-PAGE电泳分析,显示外源蛋白带在相对分子量约12kD处,以离子交换层析DEAE和分子筛纯化蛋白,缓慢复性并在C2C12细胞内检测到其蛋白活性.     

Cu2＋在凤眼莲中的积累及提取态

采用水培法和化学逐步提取法,研究水生植物凤眼莲(Eichhornia crassipes)根和叶Cu2+的累积及其提取形态.结果表明,凤眼莲中Cu2+的累积模式表现为根>叶,凤眼莲根是Cu2+主要的吸收和累积部位,具有强的吸收和累积水体环境Cu2+的能力,并且根和叶对Cu2+的累积量随培养液中Cu2+浓度的增加和处理时间的延长而上升;Cu2+在凤眼莲组织中的提取态总体表现一致为:盐酸提取态>乙醇提取态>醋酸提取态>水提取态>氯化钠提取态,盐酸提取态占优势分布,稳定且毒性低的草酸铜可能是Cu2+在凤眼莲体内的主要存在形式.盐酸提取态Cu比例的下降和乙醇提取态Cu比例的上升是凤眼莲富集器官根对Cu2+污染在提取态上的响应特征之一.     

拟穴青蟹不同形态群体ATP酶活性的季节比较

认为拟穴青蟹一对螯足均不具有腕节内刺的个体（Sp）比至少一只螯足具有腕节内刺的个体（Sr）更能适应环境温度的变化。实验比较Sp与Sr鳃ATP酶活性的季节变化,主要结果如下：Sp与Sr 鳃线粒体 ATPase 活性的季节变化模式存在较大的差异,Sr 鳃线粒体的 Na+K+-ATPase、Ca2+-ATPase和Ca2+Mg2+-ATPase活性全部表现为夏季高于冬季,Sp仅有Ca2+-ATPase活性表现为夏季高于冬季。在冬季,Sp鳃线粒体的3种ATP酶活性都显著高于Sr。总的来说,Sp对低温的代谢补偿高于Sr,从而验证Sp比Sr更能适应环境温度变化这一观点。     

人类睾丸凋亡相关基因蛋白(TSARG2)在大肠杆菌中的表达和纯化

目的获得重组TsARG2基因蛋白,为进一步研究其功能和制备特异抗体奠定基础.方法根据已报道的TsARG2基因序列,采用RT-PCR的方法获得TsARG2基因.将所得的PCR产物插入原核表达载体pQE30中,得重组质粒(pQE/TSARG2)并转化大肠杆菌(E.coli)M15,通过IPTG诱导表达出带有六个组氨酸的融合蛋白,采用镍固定金属亲和层析法纯化.结果序列分析表明TSARG2基因成熟肽编码区含有918bp,编码305个氨基酸;与GenBank(AY040204)中已报道的TSARG2分离物的核苷酸序列有100%同源性.经IPTG诱导表达,SDS-PAGE电泳分析显示,表达的融合蛋白占菌体蛋白总量的66%,分子质量约为35kDa.经Ni2 -NTAagarose纯化获得SDS-PAGE电泳下单一条带.结论在大肠杆菌中获得了TsARG2基因蛋白的高效表达,为研究其生物学功能和制备单克隆抗体奠定了基础.     

Pb^2＋污染对凤眼莲根系微生物的影响

研究系列浓度铅污染对凤眼莲根系的细菌、放线菌、霉菌和酵母菌等的活菌数和相应水样的DO（溶解氧量）、COD（化学需氧量）的影响.结果表明,Pb2＋对凤眼莲根系微生物的数量及水样DO值和COD值的影响具有时间和浓度上的双重效应.细菌为凤眼莲根系微生物的优势类群,当Pb2＋高于0.25 mmol.L-1时会减少细菌的数量,影响其净化水体的能力.DO值和COD值与凤眼莲根系微生物的数量增长存在密切关系.     

少棘蜈蚣候选杀虫肽κ-SLPTX-Ssm2b的原核表达与纯化

通过搜索NCBI数据库,获得了与具有昆虫毒性的少棘蜈蚣毒素κ-SLPTX-Ssm2a高度同源的毒素分子κ-SLPTX-Ssm2b.通过密码子优化并全基因合成获得了其DNA序列,利用RF-cloning技术将κ-SLPTXSsm2b插入到表达载体pet-HIS-SUMO,通过自动诱导表达技术在大肠杆菌BL21(DE3)诱导融合蛋白表达,通过镍柱纯化并用Ulp1激酶切割sumo标签并释放毒素分子,通过MALDI-TOF质谱鉴定其分子量为3 390.4658Da.研究结果表明获得了与理论分子量(3391.04Da)一致的κ-SLPTX-Ssm2b目的毒素分子,其表达产量为2.1mg/L,为进一步研究该候选杀虫肽分子的生理活性奠定了基础.     

白莲河水库凤眼莲过度繁殖及其防治对策

白莲河水库是鄂东最大的多功能水库．近年来凤眼莲在水库过度繁殖，已覆盖了库区水体总面积的60％．本分析了凤眼莲过度繁殖的原因并提出了防治对策．     

铅胁迫对凤眼莲氮代谢关键酶的影响

研究不同浓度Pb2＋胁迫凤眼莲0.25,6,20,60 d,其氮代谢关键酶活性的变化.结果表明：在凤眼莲根、叶中均有谷氨酰胺合成酶（GS）和谷氨酰胺α-酮戊二酸氨基转移酶（GOGAT）的分布,无论是否有Pb2＋胁迫,凤眼莲的根、叶的GS活性大于NADH-GOGAT活性,且GS活性和NADH-GOGAT活性均表现为叶大于根.0.1~2 mmol.L-1Pb2＋胁迫凤眼莲0.25,6,20,60 d,其根和叶的GS活性降低,但在胁迫60 d后随着凤眼莲新生组织的萌生,各处理浓度GS活性均出现不同程度的上升.凤眼莲叶片的GOGAT活性受Pb2＋的影响在其处理60 d后极为明显,极显著上升.     

Study on the effect of doxorubicin on expressions of genes encoding myocardial sarcoplasmic reticulum Ca2+ transport proteins and the effect of taurine on myocardial protection in rabbits

To investigate the effect of doxorubicin(DOX) on gene expression of the myocardial sarcoplasmic reticulum (SR)Ca2+ transport proteins and the mechanism of taurine(Tau) protecting cardiac muscle cells, 9 rabbits were injected with DOX , 8 rabbits with DOX and Tau, and 9 rabbits with normal saline. Cardiac function , concentration of calcium in cardiomyocytes (Myo[Ca2+]i), activity of SR Ca2+-ATPase(SERCA2a), level of SERCA2a mRNA and Ca2+ released channels(RYR2)mRNA were detected. The left ventricle tissues were observed by electron microscopy. The results showed that cardiac index, left ventricular systolic pressure, activity of SR Ca2+-ATPase and level of SERCA2a mRNA decreased , while Myo[Ca2+]i increased in DOX-treated rabbits. DOX could not affect the level of RYR2 mRNA. Tau intervention could alleviate the increase of left ventricular diastolic pressure, Myo[Ca2+]i and the decrease of SERCA2a mRNA induced by doxorubicin. The results suggested that downregulation of SERCA2a gene expression was an important mechanism of DOX-induced cardiomyopathy and that Tau could partially improve the heart function by reducing calcium overload and alleviating downregulation of SERCA2a mRNA.     

肝素酶的原核表达与表达条件优化

利用PCR技术从肝素黄杆菌克隆到肝素酶HepI基因,通过双酶切将其克隆到原核表达载体pGEX-4T-2中,转化大肠杆菌E．coliBL21感受态细胞,获得基因工程重组菌．12℃下IPTG诱导表达12h,Glutathione Sepharose 4B纯化后获得较高纯度的HepI酶蛋白．     

钙和青霉素对老化棉花种子幼苗生长的影响

研究了Ca2+和青霉素处理液对老化棉花种子幼苗生长的影响.结果表明0.03%的青霉素和0.07%的Ca2+能显著提高其过氧化物酶(POD)活性、幼苗长度、幼苗根系活力,但对幼苗叶片中叶绿素含量的影响不显著.     

Ca^2＋和Zn^2＋对甜高梁幼苗生长和部分酶活性的影响

Ca62 、Zn^2 及二混合对甜高梁进行浸种处理可促进幼苗生长,增加根长、根数及苗高;提高种子萌发过程中淀粉酶、转化酶的活性,增强呼吸速率,促进萌发,提高出苗率;SOD、POD、CAT活也有增加,提高幼苗对干旱胁迫的抵抗能力。     

Effects of fructose-1,6-diphosphate on concentration of calcium and activities of sarcoplosnic Ca^2＋-ATPase in cardiomyocytes of Adriamycin-treated rats

Objective: To observe the effects of fructose-1,6-diphosphate (FDP) on serum levels of cardiac troponin 1 (cTnl) and creatine kinase-MB (CK-MB), as well as the concentration of calcium in cardiomyocytes (Myo[Ca2 ]) and activity of sarcoplosnic Ca2 -ATPase (SRCa2 -ATPase) in Adriamycin (ADR)-treated rats. Methods: Rats were intraperitoneally injected with ADR (2.5 mg/kg every other day for 6 times) and then with different dosages of FDP (every other day for twenty-one times). Bi-antibodies sandwich Enzyme linked immune absorption assay (ELISA) was performed to detect serum level of cTnl. CK-MB was detected by monoclonal antibody, Myo[Ca2 ] was detected by fluorescent spectrophotometry and the activity of SRCa2 -ATPase was detected by inorganic phosphate method. Results: FDP (300, 600, 1200 mg/kg) significantly reduced the serum levels of cTnl and CK-MB, while at the same time decreased calcium concentration and increased SRCa2 -ATPase activity in cardiomyocytes of ADR-treated rats (P<0.01). Conclusions: FDP might alleviate the cardiotoxic effects induced by ADR through decreasing calcium level as well as increasing SRCa2 -ATPase activity in cardiomyocytes.     

Effects of combination of irbesartan and perindopril on calcineurin expression and sarcoplasmic reticulum Ca2＋-ATPase activity in rat cardiac pressure-overload hypertrophy

INTRODUCTION Left ventricular hypertrophy has been thought to be the principal predicators of predisposing risk factor of cardiac morbidity and mortality (Devereux, 1995; Levy et al., 1990). The pathogenesis that mediates cardiac hypertrophy is poorly understood. Cardiachypertrophy can be induced by hemodynamic over-load, ischemic disease, neurohumoral factors and intrinsic defects in cardiac structural protein genes (Sadoshima and Izumo, 1997; Vikstrom and Lein-wand, 1996). Another in…