烟草二脂酰甘油酰基转移酶基因(NtDGAT2)的克隆与功能分析

二脂酰甘油酰基转移酶(diacylglycerol acyltransferase,DGAT2)是植物储存油脂生物合成过程中的关键酶,对种子储存油脂累积具有重要的生理作用。本文采用电子克隆与实验相结合的方法,从烟草种子cDNA中克隆到DGAT2基因的开放阅读框序列,命名为NtDGAT2(GenBank登录号JX843807),其序列长999bp,编码332个氨基酸。多序列比对和进化分析表明该基因编码蛋白与其他植物DGAT2具有较高相似性和典型的DGAT2结构域。利用Real-time PCR定量表达分析显示Nt-DGAT2在烟草种子、花、茎、叶和根里面都有表达,且在发育中的种子和花的发育过程大量表达。酵母互补实验证实该基因编码蛋白具有DGAT酶活性。     

Acute hyperglycemia exacerbates myocardial ischemia/reperfusion injury and blunts cardioprotective effect of GIK

There is a close association between hyperglycemia and increased risk of mortality after acute myocardial infarction (AMI). However, whether acute hyperglycemia exacerbates myocardial ischemia/reperfusion (MI/R) injury remains unclear. We observed the effects of acute hyperglycemia on MI/R injury and on the cardioprotective effect of glucose-insulin-potassium (GIK). Male rats were subjected to 30 min of myocardial ischemia and 6 h of reperfusion. Rats were randomly received one of the following treatments (at 4 ml.kg(-1).h(-1) iv): Vehicle, GIK (GIK during reperfusion; glucose: 200g/l, insulin: 60 U/l, KCL: 60 mmol/l), HG (high glucose during ischemia; glucose:500 g/l), GIK + HG (HG during I and GIK during R) or GIK + wortmannin (GIK during R and wortmannin 15 min before R). Blood glucose, plasma insulin concentration and left ventricular pressure (LVP) were monitored throughout the experiments. Hyperglycemia during ischemia not only significantly increased myocardial apoptosis (23.6 +/- 1.7% vs. 18.8 +/- 1.4%, P < 0.05 vs. vehicle), increased infarct size (IS) (45.6 +/- 3.0% vs. 37.6 +/- 2.0%, P < 0.05 vs. vehicle), decreased Akt and GSK-3beta phosphorylations (0.5 +/- 0.2 and 0.6 +/- 0.1% fold of vehicle, respectively, P < 0.05 vs. vehicle) following MI/R, but almost completely blocked the cardioprotective effect afforded by GIK, as evidenced by significantly increased apoptotic index (19.1 +/- 2.0 vs. 10.3 +/- 1.2%, P < 0.01 vs. GIK), increased myocardial IS (39.2 +/- 2.8 vs. 27.2 +/- 2.1%, P < 0.01 vs. GIK), decreased Akt phosphorylation (1.1 +/- 0.1 vs. 1.7 +/- 0.2%, P < 0.01 vs. GIK) and GSK-3beta phosphorylation (1.4 +/- 0.2 vs. 2.3 +/- 0.2%, P < 0.05 vs. GIK). Hyperglycemia significantly exacerbates MI/R injury and blocks the cardioprotective effect afforded by GIK, which is, at least in part, due to hyperglycemia-induced decrease of myocardial Akt activation.     

TNFα-initiated oxidative/nitrative stress mediates cardiomyocyte apoptosis in traumatic animals

Whole body non-penetrating trauma causes myocardial infarction in humans and mechanical trauma (MT) results in cardiac dysfunction in animals. Our recent study demonstrated that incubation of cardiomyocytes with plasma isolated from MT animals causes significant cardiomyocyte apoptosis that can be blocked by neutralization of TNFα. The present study attempted to obtain direct in vivo evidence to support that overproduction of TNFα plays a causative role in trauma-induced cardiomyocyte apoptosis. Non-lethal MT caused significant TNFα overproduction (2.4-fold at 1.5 h after MT) and increased cardiomyocyte apoptosis (starting 3 h and peaking 12 h after MT). Pharmacological inhibition of TNFα with etanercept or TNFα gene deletion reduced post-trauma myocyte apoptosis (P < 0.01). Expression of iNOS and NADPH oxidase, overproduction of NO and , and excessive protein nitration in the MT heart were all significantly reduced in etanercept-treated or TNFα^−/− mice, suggesting that oxidative/nitrative stress may contribute to TNFα-initiated myocyte apoptosis in MT hearts. Additional experiments demonstrated that inhibiting iNOS (1400W) or NADPH oxidase (apocynin), or scavenging peroxynitrite (FP15) significantly reduced myocyte apoptosis in MT animals (P < 0.01). Collectively, these data demonstrated that non-lethal mechanical trauma caused significant TNFα production that in turn stimulated myocardial apoptosis via oxidative/nitrative stress.     

Identification of Differentially Expressed Genes During Anther Abortion of Taigu Genic Male Sterile Wheat by Combining Suppression Subtractive Hybridization and cDNA Array

Talgu Genlc Male Sterile Wheat （TGMSW; Trltlcum aestlvum L.）, a dominant genic male sterile germplasm, is of considerable value in the genetic Improvement of wheat because of Its stable Inherence, complete male abortion, and high cross-fertilization rate. To Identify specially transcribed genes In sterile anther, a suppression subtractlve hybridization （aSH） library was constructed with sterile anther as the tester and fertile anther as the driver. A total of 2 304 SSH Inserts amplified by polymerase chain reaction were arrayed using robotic printing. The cDNA arrays were hybridized with 32P-labeled probes prepared from the RNA of forward- and reverse-subtracted anthers. Ninety-six clones were scored as upregulated in sterile anthers compared with the corresponding fertile anthers and some clones were selected for sequencing and analysis In GenBank. Based on their putative functions, 87 non-redundant clones were classified Into the following groups： （i） eight genes Involved In metabolic processes; （11） four material transportation genes; （iii） three signal transductlon-assoclated genes; （iv） four stress response and senescence-associated protein genes; （v） seven other functional protein genes; （vi） five genes with no known function; and （vii） another 56 genes with no match to the databases. To test the hybridization efficiency, eight genes were selected and analyzed by Northern blot. The results of the present study provide a comprehensive overview of the genes and gene products Involved In anther abortion In TGMSW.     

两种新型目标分子标记技术——CDDP与PAAP

本文介绍了CDDP和PAAP这两种新型目标分子标记技术的产生原理、引物设计、PCR扩增、PCR产物检测手段、分子标记特征及带型模式,在此基础上对它们的技术特点和技术优势进行了概括,最后,对它们的应用前景进行了展望。     

Evolution of Cooperation in Spatial Traveler's Dilemma Game

Traveler''s dilemma (TD) is one of social dilemmas which has been well studied in the economics community, but it is attracted little attention in the physics community. The TD game is a two-person game. Each player can select an integer value between and () as a pure strategy. If both of them select the same value, the payoff to them will be that value. If the players select different values, say and (), then the payoff to the player who chooses the small value will be and the payoff to the other player will be . We term the player who selects a large value as the cooperator, and the one who chooses a small value as the defector. The reason is that if both of them select large values, it will result in a large total payoff. The Nash equilibrium of the TD game is to choose the smallest value . However, in previous behavioral studies, players in TD game typically select values that are much larger than , and the average selected value exhibits an inverse relationship with . To explain such anomalous behavior, in this paper, we study the evolution of cooperation in spatial traveler''s dilemma game where the players are located on a square lattice and each player plays TD games with his neighbors. Players in our model can adopt their neighbors'' strategies following two standard models of spatial game dynamics. Monte-Carlo simulation is applied to our model, and the results show that the cooperation level of the system, which is proportional to the average value of the strategies, decreases with increasing until is greater than the critical value where cooperation vanishes. Our findings indicate that spatial reciprocity promotes the evolution of cooperation in TD game and the spatial TD game model can interpret the anomalous behavior observed in previous behavioral experiments.     

利用生化及分子标记确定长穗偃麦草(Elytrigia elongatum, EE. 2n=14)染色体与小麦染色体的部分同源性

利用限制性片段长度多态性（ＲＦＬＰ）及等电聚焦（ＩＥＦ）技术确定普通小麦中国春－二倍体长穗僵麦草７个异附加系所附加的外源染色体与小麦染色体的部分同源性,共有８个生化标记,１３个ＲＦＬＰ标记在亲本间揭示了多态性。结果表明：长穗堰麦草的ＩＥ、２Ｅ、３Ｅ、４Ｅ、 ５Ｅ、６Ｅ、７Ｅ ７条染色体分别与小麦染色体的 １、２、３、４、５、６、７ ７个部分同源群具有部分同源关系,堰麦草的ＩＥ与７Ｅ、５Ｅ与７Ｅ染色体间可能发生过重排。同时,研究还分别将Ｅｓｔ－Ｅ５、Ｅｓｔ－Ｅ８位点定位于３ＥＬ,Ｐｅｒ－Ｅ１定位于７Ｅ, Ｐｅｒ－Ｅ４定位于５Ｅ,β－Ａｍｙ－Ｅ１定位于４ＥＬ染色体,并进一步将α－Ａｍｙ－Ｅ１位点定位于６Ｅ染色体长臂上。