DNA chip-based expression profile analysis indicates involvement of the phosphatidylinositol signaling pathway in multiple plant responses to hormone and abiotic treatments

The phosphatidylinositol (PI) metabolic pathway is considered critical in plant responses to many environmental factors, and previous studies have indicated the involvement of multiple PI-related gene families during cellular responses.Through a detailed analysis of the Arabidopsis thaliana genome, 82 polypeptides were identified as being involved in PI signaling. These could be grouped into different families including PI synthases (PIS), PI-phosphate kinases (PIPK),phospholipases (PL), inositol polyphosphate phosphatases (IPPase), inositol polyphosphate kinases (IPK), PI transfer proteins and putative inositol polyphosphate receptors. The presence of more than 10 isoforms of PIPK, PLC, PLD and IPPase suggested that these genes might be differentially expressed during plant cellular responses or growth and development. Accordingly, DNA chip technology was employed to study the expression patterns of various isoforms.In total, 79 mRNA clones were amplified and used for DNA chip generation. Expression profile analysis was performed using samples that represented multiple tissues or cellular responses. Tested samples included normal leaf, stem and flower tissues, and leaves from plants treated with various hormones (auxin, cytokinin, gibberellin, abscisic acid and brassinosteroid) or environmental factors (temperature, calcium, sodium, drought, salicylic acid and jasmonic acid).Results showed that many PI pathway-related genes were differentially expressed under these experimental conditions.In particular, the different isoforms of each family were specifically expressed in many cases, suggesting their involvement in tissue specificity and cellular responses to environmental conditions. This work provides a starting point for functional studies of the relevant PI-related proteins and may help shed light onto the role of PI pathways in development and cellular responses.     

PCR-RFLP检测LDL受体基因TaqⅠ多态性位点的研究

应用聚合酶链反应(PCR)扩增人类LDL受体基因外显子4-内含子4-外显子5片段,PCR产物为1.55kb,DNA片段经序列鉴定后,进行TaqI酶切位点的RFLP分析。结果显示:中国汉族人群LDL受体基因中存在着TaqⅠ酶切位点多态性; 200个LDL受体等位基因中TaqⅠ酶切位点出现的频率为0.515,该点频率较为适中, 可作为中国汉族人群LDL受体基 因的遗传标志来进行家族性高胆固醇血症(FH)的基因诊断。所建立起的LDL受体基因TaqⅠ位点的PCR -RFLP方法具有快速、简便的特点,在FH的基因诊断上有应用价值。 Abstract:To develop rapid and sensitive technique for detectin the TaqI polymorphism at the human LDL receptor gene in Chinese,the exon4-intron4-exon5 of the human LDL receptor gene was amplified by polymerase chain reaction(PCR).The PCR products were directly analysed by restriction fragment length polymorphisms(RFLP).The results showed that the TaqI polymorphism is associated with the LDL receptor gene in Chinese of Han nationality;The frequency of T= allele (presence of TaqI cutting site)is 0.515 in 200 LDL receptor alleles.This technique may be used for rapid and sensitive screening of the LDL receptor gene for the TaqI polymorphism.     

浙江地方猪种线粒体DNA多态及遗传多样性研究

本实验用ＡｐａＩ、ＡｖａＩ、ＢａｍＨⅠ、ＢｃｌⅠ、ＢｇｌⅠ、ＢｇｌⅡ、ＣｌａⅠ、ＤｒａⅠ、Ｅｃｏ ＲＩ、Ｅｃｏ ＲＶ、Ｈａｅ Ⅱ、Ｈｉｎｃ Ⅱ、ＨｉｎｄⅢ、ＨｐａⅠ、ＫｐｎⅠ、ＰｖｕⅠ、ＰｖｕⅡ、Ｓａｃ Ⅰ、ＳａｌⅠ、ＳｃａⅠ、ＳｍａⅠ、ＳｔｕⅠ、ＸｂａⅠ和ＸｈｏⅠ等２５种识别６个碱基对的限制性内切酶分析了浙江地方品种猪、浙江野猪等３０个个体的线粒体ＤＮＡ,共检出３０种限制性态型,可归结成３种单倍     

血清CA199、CYFR21-1、CEA、NSE水平与肺癌的相关性研究

目的：分析血清CA199、CYFR21-1、CEA、NSE 水平检测在肺癌诊断及疗效评估中的临床价值,探讨其与肺癌的相关性。方 法：采用化学发光免疫法检测78 例肺癌患者、50 例肺部良性疾病及50 例健康体检者血清CA199、CYFR21-1、CEA、NSE 水平,比 较治疗前后肺癌患者肿瘤标志物水平的变化。结果：肺癌组血清CA199、CYFR21-1、CEA、NSE 水平较对照组、肺良性疾病组均显 著升高(P<0.05);Ⅲ-Ⅳ期肺癌患者各肿瘤标志物水平均较Ⅰ-Ⅱ期患者明显升高（P<0.05）;四项联合检测的敏感性为79.5 %,明显 高于CA199、CYFR21-1、CEA、NSE 任一单项肿瘤标志物的敏感性（P<0.05）;随着疗效的下降,血清CA199、CYFR21-1、CEA、NSE 水平逐渐升高（P<0.05）。结论：肺癌患者血清CA199、CYFR21-1、CEA、NSE 水平呈高表达状态,且其表达水平与肺癌病情程度密 切相关,联合检测有助于提高肺癌诊断的敏感度及指导治疗方案选择。     

Polymorphisms in PPARD, PPARG and APM1 associated with four types of Traditional Chinese Medicine constitutions

Based on the theory of constitution of Traditional Chinese Medicine (TCM), the human population is divided into nine constitutions including one balanced constitution (Normality) and eight unbalanced constitutions (Yang-deficiency, Yin-deficiency, Phlegm-wetness, Qi-deficiency, Wetness-heat, Blood stasis, Depressed constitution, and Inherited special constitution). Different constitutions have specific metabolic features and different susceptibility to certain diseases. However, whether a genetic basis accounts for such constitution classification is yet to be determined. Here we performed a genetic study to assess the association between genetic variations of metabolic genes including PPARD, PPARG and APM1 and the constitutions. A total of 233 individuals of the Han population in China were classified into four groups, Normality, Yang-deficiency, Yin-deficiency and Phlegm-wetness with whom 23 single nucleotide polymorphisms (SNPs) in the three genes were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Biased distribution of PPARD rs2267669 and rs2076167, APM1 rs7627128 and rs1063539 in Yang-deficiency, PPARG Pro12Ala in Yin-deficiency and PPARD rs2076167, APM1 rs266729 and rs7627128 in Phlegm-wetness were observed. The frequencies of Haplotype13 (Hap13) of PPARG in Yin-deficiency, Hap25 of APM1 in Yang-deficiency and Hap2 of PPARD and Hap14 of PPARG in Phlegm-wetness, were significantly different from those in Normality, suggesting those might be group-associated haplotypes. These results suggested that single SNP and haplotypes of PPARD, PPARG and APM1 may underlie the genetic basis of the constitutions classified in TCM.     

从猕猴肝脏组织扩增黄嘌呤脱氢酶／氧化酶基因片段

RT-PCR扩增猕猴黄嘌呤脱氢酶／氧化酶（XDH／XO）基因片段,为进一步开展相关研究提供实验资料。方法提取猕猴新鲜肝脏组织总RNA,用RT-PCR二步法进行XDH／XO基因片段扩增,对获得的目的片段进行序列测定,与GenBank上发表的人类（Homosapiens）、小鼠（Musmusculus）、家鼠（Rattusnorvegicus）、野猪（Susscrofa）等物种XDH／XO基因进行该序列同源性比对分析,DNAMAN软件预测该段核苷酸的氨基酸序列,Inter-ProScan及SWISS-MODEL工具进行XDH／XO的编码蛋白结构域及功能预测及三维结构构建。结果RT-PCR产物电泳检测得到了与设计大小相一致的目的条带,序列测定共测到683个核苷酸,DNAMAN软件预测该段核苷酸的氨基酸序列包括了1个编码53个氨基酸的开放阅读框（ORF）,通过该软件包中Multiplealignment对目的基因片段的核苷酸序列与NCBI报道的人类、小鼠、家鼠、野猪XDH／XO基因mRNA互补的cDNA核苷酸序列同源性进行同源性比较分析,结果显示所扩增得到的目的片段与人类同源性最高,为95．6％,与小鼠、家鼠、野猪的同源性分别为85．2％、84．3％、86．1％,说明获得的基因片段是猕猴的XDH／XO基因片段,且该基因在物种间具有较高的相似性。生物信息学预测该段XDH／XO编码蛋白含有醛氧化／脱氢酶的钼喋呤结合点结构域及黄嘌呤脱氢酶结构域。结论在体外成功扩增出猕猴XDH／XO基因片段,为进一步开展高尿酸血症致病机理研究,抗高尿酸血症新药研发奠定工作基础。     

西双版纳傣族“龙山”片断热带雨林蚂蚁类群结构与多样性研究

在西双版纳热带雨林地区,采用陷阱法（Pitfall traps) 对2类龙山片断雨林和保护区连续雨林蚂蚁类群进行取样调查,样地内共获取蚂蚁9196号,隶属6亚科,28属,53种,其属种和个数数量以切叶蚁亚科（Myrmicinae)最多。不同生境群落组成分差异较大,优势种变化明显。对3种蚂蚁类群多样性研究分析表明：保护区连续雨林的多样性、均匀性指数高于龙山片断林,群落相似性以2类龙山林最高,各林地蚂蚁类群物种分布的种－多度关系为对数级数模式。片断化雨林面积、植被结构、隔离程度及状况和人为干扰程度是影响蚂蚁类群结构和多样性的重要因素。     

基因检测技术 探针在诊断中的应用

在过去几十年来,世界上用分子水平分析的方法来研究人的基因失调已有许多报告,增加了很多新的知识,这些知识是由于生化的重组基因技术而来,用在单独的分析特异的基因、基因产物以及核酸结构分析编码,基因表达的规律,从而用以检测某些疾病,例如早期妊娠时测定血清或羊水中的基因紊乱,可以及时治疗或预防。     

三倍体白杨杂种无性系指纹图谱的构建

目的:构建三倍体白杨杂种无性系指纹图谱,鉴定三倍体白杨杂种无性系。方法:分离纯化三倍体白杨杂种DNA模板,采用扩增片段长度多态性(AFLP)分子标记技术构建三倍体白杨杂种无性系指纹图谱。结果:从64对引物组合中筛选出M-CTA/E-CAG、M-CAC/E-CCA、M-ACT/E-CTC和M-CTT/E-CTG等4对多态性较高的引物组合,并应用该引物组合对21个三倍体白杨杂种无性系进行了AFLP分析,构建了21个三倍体白杨杂种无性系指纹图谱。结论:构建无性系指纹图谱是鉴别三倍体白杨杂种无性系的有效方法,能够有效鉴别21个三倍体白杨杂种无性系。本研究为品种鉴定及新品种权保护奠定了基础。     

水稻土中铁还原菌多样性

微生物介导的异化Fe(III) 还原是非硫厌氧环境中Fe(III) 还原生成Fe(II) 的主要途径,然而相关的铁还原菌还不是很清楚,特别是在水稻土中.本文采用富集培养的方法,以乙酸和氢气作为电子供体,水铁矿和针铁矿作为电子受体,通过末端限制性片段长度多态性（T-RFLP）技术和16S rRNA基因克隆测序相结合的分子生物学方法研究了水稻土中铁还原菌的多样性.结果表明：无论是以乙酸或氢气为电子供体,水铁矿或针铁矿为电子受体,地杆菌（Geobacter）和梭菌（Clostridiales）是富集到的主要微生物群落;乙酸为电子供体时,富集到的主要微生物群落还包括红环菌（Rhodocyclaceae）;因此,除地杆菌外,梭菌和红环菌很可能也是水稻土中重要的铁还原菌.