陆地棉一个丝氨酸/苏氨酸激酶蛋白基因的克隆与表达分析   总被引：3，自引：1，他引：3  

杨郁文  倪万潮  张保龙  沈新莲  张香桂  徐英俊  姚姝 《棉花学报》2006,18(3):140-144

通过RT-PCR获得一个编码棉花丝氨酸/苏氨酸激酶类似蛋白全长的cDNA片段,其编码的氨基酸序列与拟南芥ATPK3的相似度达到82%,这个基因暂被命名为GhPK1。基因全编码区包括1038个核苷酸,编码346个氨基酸的蛋白。GhPK1的编码产物在265~270个氨基酸处有一个跨膜区并可能结合在内质网膜上。GhPK1在种子和纤维中的表达水平较其它组织高。另外,GhPK1的表达量在受到盐胁迫后上升,说明GhPK1可能参与了盐胁迫反应。  相似文献   

Quercetin activates mitochondrial autophagy via PINK1/parkin pathway to alleviate cerebral ischemia-reperfusion injury in rats     

WEI Si-can  LIN Tian-lai  HUANG Ling  SU Zhi-wei  WEI Mei-juan 《园艺学报》2021,36(12):2251-2257

AIM To analyze the regulatory effect of quercetin （QUE） on PTEN-induced putative kinase 1 （PINK1）/parkin mitochondrial autophagy pathway, and to explore the mechanism of quercetin in relieving cerebral ischemia/reperfusion （I/R） injury. METHODS Sixty SD male rats were randomly divided into sham operation group, model group （I/R group）, QUE group,3-methyladenine （3-MA） group and QUE+3-MA group. Administration started in each group 3 days before modeling, once a day, at 30 min after the last administration,except sham group, the other groups used 4-vessel blockage method to establish the whole brain I/R model. On the day after modeling, the neural function was evaluated by neuropathy disability score （NDS）. The volume of cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride （TTC） staining. The morphological changes of mitochondria in hippocampus were observed by transmission electron microscopy. The contents of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in hippocampus were measured by ELISA. The activity of superoxide dismutase （SOD） and contents of malondialdehyde （MDA） in hippocampus were detected by xanthine oxidase method, thiobarbituric acid condensation method. Western blot was used to detect the proteinex pression of PINK1, parkin and LC3-II in brain tissue. RESULTS Compared with sham group, the hippocampus of the rats in I/R group and QUE+3-MA group showed swelling of mitochondria, destruction or disappearance of internal crista and other pathological damage,also the volume of cerebral infarction, the contents of IL-6, TNF-α and MDA, the protein expression levels of PINK1, parkin and LC3-II were increased （P<0.05）, while NDS score and activity of SOD were decreased （P<0.05）. Compared with I/R group and QUE+3-MA group, the pathological damage degree of hippocampus in QUE group was reduced, the volume of cerebral infarction, the contents of IL-6, TNF-α and MDA were decreased （P<0.05）, the proteinexpression levels of PINK1, parkin and LC3-II, and NDS score and activity of SOD were increased （P<0.05）.The above indexes in 3-MA group were opposite to QUE group. No significant difference in the above indexes between I/R group and QUE+3-MA group was observed （P>0.05）. CONCLUSION Quercetin activates mitochondrial autophagy and reduces cerebral I/R by regulating the expression of PINK1/parkin pathway proteins.  相似文献   

编码杜果丝氨酸/苏氨酸蛋白激酶类蛋白抗病基因同源序列的克隆与分析     

张玄兵  杜中军  黄俊生  王家保  徐兵强 《华中农业大学学报》2006,25(2):186-189

根据丝氨酸／苏氨酸蛋白激酶类催化结构域Ⅰ和Ⅸ的氨基酸保守序列设计简并引物,PCR扩增‘紫花芒’杧果嫩绿色叶片基因组DNA。在NCBI中用Blast X比对发现,有6个阳性克隆是抗病基因同源序列,并命名为PK1～PK6,GenBank注册序列号为AY693369-AY693371和AY776277-AY776279。在PK3、PK4和PK6中发现了丝氨酸／苏氨酸蛋白激酶的9个催化结构域Ⅰ～Ⅸ,而其它3个克隆具有部分结构域。氨基酸序列同源性分析还发现,6个克隆编码的氨基酸序列都与受体样蛋白激酶有较高的氨基酸序列同源性。系统进化树分析还表明它们都是可能的抗病基因同源序列。总之,6个克隆不仅是抗病基因同源序列而且是可能的受体样蛋白激酶基因同源序列。  相似文献   

Silencing arginine kinase/integrin β1 subunit by transgenic plant expressing dsRNA inhibits the development and survival of Plutella xylostella     

Shu Fu  Zhaoxia Liu  Jinzhi Chen  Gengxiao Sun  Yingxia Jiang  Miaowen Li  Lei Xiong  Shaoping Chen  Yuqing Zhou  Muhammad Asad  Guang Yang 《Pest management science》2020,76(5):1761-1771

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ROCK promotes high glucose-induced cardiomyocyte apoptosis by inhi-biting PI3K/Akt signaling pathway     

BAI Xue  YU Hui  ZHU Ju-ru  HAN Zhong-li  YANG Xiao-min 《园艺学报》2020,36(5):788-795

AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway, thus promoting the apoptosis of cardiomyocytes.  相似文献   

小麦属Xa 21-类似蛋白激酶基因的克隆及与同源位点序列比较     

牛吉山  刘瑞  王林海  郑磊 《河南农业大学学报》2007,41(5):475-479

对小麦属Xa21-类似蛋白激酶基因进行了克隆及与同源序列比较研究.在小麦(Triticum aestivum)高分子量麦谷蛋白基因位点附近有一个编码LRR-类受体蛋白激酶的基因位点(暂标记为TaXa),编码蛋白与水稻(O ryza sativa)抗病蛋白Xa 21类似.通过反转录PCR途径,从普通小麦和二粒小麦(Triticum turgidum)的TaXa同源位点分离了3个cDNA克隆,ZS 860(GenBank查询号:EF 394367),ZS 2000(GenBank查询号:EF 394368)和ZS 2001(GenBank查询号:EF 394369).TaXa位点的祖先基因可能编码1 028个氨基酸组成的多肽.一个完整的TaXa蛋白包括N-端保守区、LRR结构域、一个跨膜区和位于C—端的丝氨酸—苏氨酸蛋白激酶功能域.在基因进化过程中,由于碱基代换、缺失和插入导致了开放读码框的改变,使该位点基因的编码多肽缩短.本研究对小麦属TaXa同源位点编码氨基酸序列和1个大麦(Hordeum vulgare)中的同源蛋白进行了详细比较.  相似文献   

鱼类精氨酸需求研究进展   总被引：1，自引：2，他引：1  

罗智 《水产学报》2004,28(4):450-459

精氨酸是迄今研究的所有鱼类的必需氨基酸。近年来,在确定鱼类精氨酸需要量方面取得了巨大的进展。然而,仍然有许多养殖鱼类对精氨酸的需求有待研究。在已有的报道中,鱼类精氨酸需求存在种内和种间差异,而且这种差异较大。这篇论文综述了鱼类精氨酸需求的研究进展,包括五方面的内容,即鱼类对精氨酸的需要量,饲料中精氨酸含量对鱼类的影响,影响鱼类精氨酸需要量的因素,评价鱼类精氨酸需要量所采用的指标以及精氨酸和赖氨酸之间的相互关系。  相似文献   

大菱鲆MKK基因家族全基因组鉴定及其在生物和非生物应激下的表达分析     

郑卫卫  徐锡文  陈松林  俄泽琛  刘英杰 《水产学报》2023,47(1):019109-019109

为了阐明大菱鲆丝裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinases,MAPKKs或MKKs)基因家族在生物和非生物应激响应中的作用,本实验首先通过生物信息学方法对大菱鲆MKK基因家族进行了全基因组水平的鉴定,利用多个应激相关转录组数据集分析了大菱鲆MKK家族成员在不同组织及不同生物和非生物应激下的表达模式。结果显示,本研究在大菱鲆全基因组水平上共鉴定出9个MKK基因家族成员,它们不均匀地分布在7条染色体上,并分别对其编码蛋白的理化性质、蛋白二级结构和亚细胞定位进行了预测。基于系统发育分析,将SmMKKs划分为5个亚家族。内含-外显子结构、保守基序和多重序列比对分析结果不仅为大菱鲆MKK亚家族分类提供了证据,而且表明SmMKKs在进化上高度保守。SmMKKs在不同组织及不同生物和非生物应激下的基因表达模式分析表明,SmMKKs具有明显的组织特异性表达。另外,结果显示,粘孢子虫和肿大细胞病毒感染后,SmMKK6a呈极显著差异表达;热应激处理后,SmMKK6a呈极显著差异表达;高盐或低盐胁迫后,SmMKK4a、SmMKK4b、SmMKK6a...  相似文献   

Effects of different arginine/lysine level on growth performance,body composition and digestive enzyme activity of Macrobrachium rosenbergii          下载免费PDF全文

X. J. Dong  J. Wu  Y. Shen  J. Y. Chen  S. Y. Miao  X. J. Zhang  L. S. Sun 《Aquaculture Nutrition》2018,24(3):1101-1111

Given the universal presence of antagonism between arginine and lysine, determining the optimal dietary arginine/lysine level is of great importance. In this study, eight diets were formulated to contain different arginine/lysine level, followed by a 9‐week feeding trial with shrimps of 2.10 ± 0.01 cm in length and 0.38 ± 0.02 g in weight to determine the effect of dietary arginine/lysine level on growth performance, body composition and enzyme activity. The results showed that the Diet 6 (arginine/lysine level of 29.4/21.0 g/kg) group had significantly higher weight gain rate, and specific growth rate than the control group and most of the other groups (p < .05) did. Similarly, the retention efficiency of arginine and lysine in the Diet 6 group was significantly higher than the control group and most of the other groups (p < .05). Moreover, most digestive enzymes had higher activity levels in the Diet 6 group, and evident antagonism between dietary arginine and lysine was detected. In summary, dietary arginine/lysine level of 29.4/21.0 g/kg produced the best growth performance, suggesting that a better growth performance in Macrobrachium rosenbergii culture can be obtained by adjusting the dietary arginine/lysine level.  相似文献   

甘蓝自交不亲和决定因子的体外表达和相互作用的检测   总被引：5，自引：3，他引：2  

杨洋  高启国  宋明  牛义  汤青林  朱利泉  王小佳 《园艺学报》2009,36(3):355-362

 S位点受体激酶(SRK)和S位点富含半胱氨酸蛋白/S位点蛋白11(SCR/SP11)分别为甘蓝自交不亲和(self-incompatibility, SI)信号传导的雌雄决定因子。为了深入研究两者的作用机理和进行人工调控,本研究以结球甘蓝ZQ为材料,利用pET·NusA融合蛋白表达系统,将包含SRK胞外域和跨膜域的mSRK蛋白和SCR蛋白在大肠杆菌BL21中融合表达,经SDS-PAGE电泳检测表达出融合蛋白大小分别约为116 kD和74 kD。进一步将两融合蛋白进行体外相互作用的检测,结果表明mSRK与SCR蛋白能够相互结合形成稳定的复合体,这为下一步实现SRK-SCR复合体聚合与解离的人为调控提供了技术平台。  相似文献