一个玉米类ABC1基因ZmABC1-10的克隆及其对镉等非生物胁迫的应答(英文)

镉是一种非必需的重金属元素,对动植物有严重毒害作用。几个与ABC1(activity of the bc1 complex)家族有关的基因参与植物镉胁迫的应答。本研究从玉米中克隆并鉴定了一个类ABC1基因,命名为ZmABC1-10。该基因cDNA全长2 519 bp,包含一个2 250 bp的开放阅读框,编码一个预测的叶绿体膜蛋白。启动子顺式元件扫描发现该基因含有大量的非生物胁迫、光以及植物激素应答元件。表达模式分析表明,该基因主要在叶片、茎秆等绿色组织中表达。镉处理实验表明,该基因能够被诱导并且受植物发育时期的调控。除镉之外,该基因还受多种非生物因素包括ABA、H2O2、干旱和黑暗的共同调控。此外,本研究利用基因组序列信息共鉴定出19个玉米ABC1基因。对植物界8个代表性物种中148个ABC1蛋白进行系统发育分析表明,在长期进化过程中植物ABC1蛋白已经发生了分化;物种特异性扩张是植物中该家族进化的主要动力。这些结果表明ZmAbc1-10是一个镉应答因子并且可能在植物对非生物胁迫的适应中发挥重要作用。

ABCE1基因siRNA表达质粒的构建及鉴定

目的:构建针对人ABCE1基因siRNA表达质粒,为进一步研究该基因功能奠定基础。方法:合成含靶向ABCE1基因siRNA转录模板的茎环结构,与两端分别有BamHⅠ、HindⅢ酶切位点的RNAi-Ready pSIREN-DNR-DsRed-Express质粒连接,大肠杆菌中扩增,测序鉴定。结果:转化大肠杆菌涂布平板长出阳性菌落,重组质粒电泳初步说明质粒构建成功,测序结果表明RNAi-Ready pSIREN-DNR-DsRed-Express的酶切位点BamHⅠ、HindⅢ之间有71bp的插入片段,其序列与所设计、合成的ABCE1的siRNA转录模板序列一致。结论:siRNA转录模板完整正确地插入RNAi-Ready pSIREN-DNR-DsRed-Express质粒中。

烟草氧化胁迫相关基因NtOSA1的克隆表达及亚细胞定位分析

植物类蛋白激酶Abc1家族(activity of bc1complex)在植物生长发育及响应非生物胁迫中起着重要的作用。该研究通过将拟南芥AtOSA1蛋白氨基酸序列与烟草转录组数据进行比对,利用巢式PCR技术克隆得到1个烟草氧化胁迫相关Abc1家族基因NtOSA1,NtOSA1与拟南芥AtOSA1基因具有72.89%的一致性。NtOSA1基因开放阅读框长度为2 283bp,编码760个氨基酸,含有典型的ABC1结构域、一个激酶结构域、叶绿体定位信号肽和两个跨膜结构域。采用实时荧光定量PCR技术,对NtOSA1基因在烟草不同组织以及氧化胁迫、盐胁迫等处理下的表达分析表明:NtOSA1基因的表达具有组织特异性,主要在叶片中表达;NtOSA1基因在H2O2和NaCl处理后表达量上升,均在处理后6h达到最大值,分别为处理前的1.95和2.69倍。亚细胞定位结果表明,NtOSA1蛋白定位在细胞叶绿体上,与预测结果一致。研究表明,NtOSA1基因参与了烟草抗氧化胁迫和盐胁迫的响应。

Genome-wide identification of abscisic acid(ABA) receptor pyrabactin resistance 1-like protein(PYL) family members and expression analysis of PYL genes in response to different concentrations of ABA stress in Glycyrrhiza uralensis

As abscisic acid(ABA)receptor,the pyrabactin resistance 1-like(PYR/PYL)protein(named PYL for simplicity)plays an important part to unveil the signal transduction of ABA and its regulatory mechanisms.Glycyrrhiza uralensis,a drought-tolerant medicinal plant,is a good model for the mechanism analysis of ABA response and active compound biosynthesis.However,knowledge about PYL family in G.uralensis remains largely unknown.Here,10 PYLs were identified in G.uralensis genome.Characterization analysis indicated that PYLs in G.uralensis(Gu PYLs)are relatively conserved.Phylogenetic analysis showed that Gu PYL1-3 belongs to subfamily I,Gu PYL4-6 and Gu PYL10 belong to subfamily II and Gu PYL7-9 belongs to subfamily III.In addition,transcriptome data presented various expression levels of Gu PYLs under different exogenous ABA stresses.The expression pattern of Gu PYLs was verified by Quantitative real-time polymerase chain reaction(q RT-PCR).The study proved that Gu PYL4,Gu PYL5,Gu PYL8 and Gu PYL9 genes are significantly up-regulated by ABA stress and the response process is dynamic.This study paves the way for elucidating the regulation mechanism of ABA signal to secondary metabolites and improving the cultivation and quality of G.uralensis using agricultural strategies.

乳腺癌组织及淋巴结中细胞间粘附分子表达与其浸润转移关系探讨

目的探讨细胞间粘附分子（ICAM-1）表达与乳腺癌的关系。方法应用免疫组织化学ABC法检测乳腺癌及其腋下淋巴结中ICAM－1的表达．结果乳腺癌组织中ICAM-1的表达明显高于乳腺腺癌，转移癌与未转移癌组之间相比差异非常显著，且与病理分级呈负相关趋势，腋下转移淋巴结与非转移淋巴结中ICAM-1表达阳性率无差别，但阳性度方面明显不同。结论乳腺癌组织中ICAM－1检测可以作为判定患者机体免疫功能状态及预后的客观指标．

Interaction of the major inflammatory bowel disease susceptibility alleles in Crohn’s disease patients

AIM:To investigate the interaction of interleukin-23 receptor(IL23R)(rs1004819 and rs2201841),autophagy-related 16-like 1(ATG16L1)(rs2241880), caspase recruitment domain-containing protein 15 (CARD15)genes,and IBD5 locus in Crohn's disease(CD) patients. METHODS:A total of 315 unrelated subjects with CD and 314 healthy controls were genotyped.Interactions and specific genotype combinations of a total of eight variants were tested.The variants of IBD5locus(IGR2198a_1 rs11739135 and IGR2096a_1 rs12521868),CARD15(R702W rs2066845 and L1007fs rs2066847),ATG16L1(rs2241880)and IL23R (rs1004819,rs2201841)genes were genotyped by PCR-RFLP,the G908R(rs2066844)in CARD15 was determined by direct sequencing. RESULTS:The association of ATG16L1 T300A with CD was confirmed[P=0.004,odds ratio(OR)=1.69, 95%CI:1.19-2.41],and both IL23R variants were found to represent significant risk for the disease(P= 0.008,OR=2.05,95%CI:1.20-3.50 for rs1004819 AA;P<0.001,OR=2.97,95%CI:1.65-5.33 for rs2201841 CC).Logistic regression analysis of pairwise interaction of the inflammatory bowel disease (IBD)loci indicated that IL23R,ATG16L1,CARD15 and IBD5(IGR2198a_1)contribute independently to disease risk.We also analysed the specific combina- tions by pair of individual ATG16L1,IL23R rs1004819, rs2201841,IGR2198a_1,IGR2096a_1 and CARD15 genotypes for disease risk influence.In almost all cases,the combined risk of susceptibility pairs was higher in patients carrying two different risk-associated gene variants together than individuals with just one polymorphism.The highest OR was found for IL23R rs2201841 homozygous genotype with combination of positive CARD15 status(P<0.001,OR=9.15,95% CI:2.05-40.74). CONCLUSION:The present study suggests a cumulative effect of individual IBD susceptibility loci.