The mRNA Expression Profiles of Five Heat Shock Protein Genes from Frankliniella occidentalis at Different Stages and Their Responses to Temperatures and Insecticides

The westem flower thrips, Frankliniella occidental＆ （Pergande） is a highly invasive pest that is able to exploit many crops across a wide range of environmental conditions. Five full-length cDNAs of heat shock protein （HSP） genes （Fo-HSP90, Fo-HSP70, Fo-HSP60, Fo-HSP40 and Fo-HSP28.9） were cloned from F. occidentalis, and their expression profiles were investigated under conditions of thermal stress and insecticide exposure, and at different stages during development, using real-time quantitative PCR. All five gene sequences showed high similarity to homologs in other species, indicating the conserved fimction of this gene family. HSP60 represents an informative phylogenetic marker at the ordinal taxonomic level within Insecta, but HSP90, which has two homologous copies in Hymenoptera, was not informative. The expression of Fo-HSPs under thermal stress suggests that Fo-HSP90, Fo-HSP70, and Fo-HSP28.9 are inducible by both cold and heat stress, Fo-HSP40 is only heat-inducible, and Fo-HSP60 is thermally insensitive. There were two patterns of cold induction of Fo-HSPs： one is from 0 to 4℃ and the other is around -8℃. All five Fo-HSPs genes were induced by exposure to sublethal concentrations of the insecticide avermectin. The expression of the five Fo-HSPs during different developmental stages suggests that they all play a role in development of F. occidentalis.

Genomic characterization and expression profiles of stress-associated proteins(SAPs)in castor bean(Ricinus communis)

Stress-associated proteins(SAPs)are known as response factors to multiple abiotic and biotic stresses in plants.However,the potential physiological and molecular functions of SAPs remain largely unclear.Castor bean(Ricinus communis L.)is one of the most economically valuable non-edible woody oilseed crops,able to be widely cultivated in marginal lands worldwide because of its broad adaptive capacity to soil and climate conditions.Whether SAPs in castor bean plays a key role in adapting diverse soil conditions and stresses remains unknown.In this study,we used the castor bean genome to identify and characterize nine castor bean SAP genes(RcSAP).Structural analysis showed that castor bean SAP gene structures and functional domain types vary greatly,differing in intron number,protein sequence,and functional domain type.Notably,the AN1-C2H2eC2H2 zinc finger domain within RcSAP9 has not been often observed in other plant families.High throughput RNA-seq data showed that castor bean SAP gene profiles varied among different tissues.In addition,castor bean SAP gene expression varied in response to different stresses,including salt,drought,heat,cold and ABA and MeJA,suggesting that the transcriptional regulation of castor bean SAP genes might operate independently of each other,and at least partially independent from ABA and MeJA signal pathways.Cis-element analyses for each castor bean SAP gene showed that no common cis-elements are shared across the nine castor bean SAP genes.Castor bean SAPs were localized to different regions of cells,including the cytoplasm,nucleus,and cytomembrane.This study provides a comprehensive profile of castor bean SAP genes that advances our understanding of their potential physiological and molecular functions in regulating growth and development and their responses to different abiotic stresses.

Genome-wide characterization of mapk gene family in black rockfish Sebastes schlegelii and their expression patterns against Edwardsiella piscicida infection

Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.

Discovery and validation of prognostic markers in gastric cancer by genome-wide expression profiling

AIM: To develop a prognostic gene set that can predict patient overall survival status based on the whole genome expression analysis. METHODS: Using Illumina HumanWG-6 BeadChip followed by semi-supervised analysis, we analyzed the expression of 47 296 transcripts in two batches of gastric cancer patients who underwent surgical resection. Thirty-nine samples in the first batch were used as the training set to discover candidate markers correlated to overall survival, and thirty-three samples in the second batch were used for validation. RESULTS: A panel of ten genes were identified as prognostic marker in the first batch samples and classified patients into a lowand a high-risk group with significantly different survival times (P = 0.000047). This prognostic marker was then verified in an independent validation sample batch (P = 0.0009). By comparing with the traditional Tumor-node-metastasis (TNM) staging system, this ten-gene prognostic marker showed consistent prognosis results. It was the only independent prognostic value by multivariate Cox regression analysis (P = 0.007). Interestingly, six of these ten genes are ribosomal proteins, suggesting a possible association between the deregulation of ribosome related gene expression and the poor prognosis. CONCLUSION: A ten-gene marker correlated with overall prognosis, including 6 ribosomal proteins, was identified and verified, which may complement the predictive value of TNM staging system.

Genome-wide identification,phylogeny and expression analysis of the SBP-box gene family in maize(Zea mays)

The SQUAMOSA promoter binding protein （SBP）-box genes encode a kind of plant-specific transcription factors （TFs） and play important roles in the regulation of plant development. In this study, a genome-wide characterization of this family was conducted in maize （Zea mays）. Thirty-one SBP-box genes were identified to be distributed in nine chromosomes and 16 of them were complementary to the mature ZmmiR156 sequences. All the Z. mays SBP （ZmSBP） genes were classified into two clusters with eight subgroups according to the phylogenetic analysis of proteins, which were consistent with the pattern of exon-intron structures. The phylogenetic tree of the ZmSBP, Oryza sativa SBP-like （OsSPL） and Arabidopsis thaliana SBP-like （AtSPL） genes were constructed and all the SBP-box genes were divided into eight groups, which was the same as the classification of ZmSBP genes. The comparision of the expression profiles of all SBP-box genes in these three species indicated that most orthologous genes had similar expression patterns. The results from this study provided a basic understanding of the ZmSBP genes and might facilitate future researches for elucidating the SBP-box genes function in maize.

Differential Expression of Rice Valine-Qlutamine Gene Family in Response to Nitric Oxide and Regulatory Circuit of OsVQ7 and OsWRKY24

The functional diversity of plant valine-qlutamine(VQ) proteins is closely associated with their partners WRKY transcription factors, and also with a complex network of signaling pathways that mediated by hormone molecules. We reported genome-wide expression profiles of differentially expressed rice VQ genes under nitric oxide(NO) treatment based on a microarray analysis. Cluster analysis of expression patterns revealed that some VQ genes and WRKY genes shared similar expression trends. Prediction of cis-elements showed that W-box or W-box-like sequences were overrepresented within the promoters of most of NO-responsive VQ genes. In particular, the similarly expressed Os VQ7 and Os WRKY24 showed great induction upon NO triggering. Transient expression assay and chromatin immunoprecipitation analysis demonstrated that OsWRKY24 was specifically bound to the promoter regions of Os VQ7 and Os WRKY24 itself, which contain multiple copies of W-box or W-box-like cis-elements. Yeast-two-hybrid assay indicated that OsWRKY24 can interact physically with OsVQ7 through the C-terminal of WRKY domain. The results suggested that OsVQ7 and OsWRKY24 may form an auto-and cross-regulation circuit that is required for tight regulation and fine-tuning of physiological processes they are involved in. These findings provided a solid foundation for exploring the specific functions of the VQ protein family in NO signaling pathway.

Characterization of the chemosensory protein EforCSP3 and its potential involvement in host location by Encarsia formosa

Chemosensory proteins(CSPs) perform several functions in insects.This study performed the gene expression,ligand-binding,and molecular docking assays on the EforCSP3 identified in the parasitoid wasp Encarsia formosa,to determine whether EforCSP3 functions in olfaction,especially in host location and host preference.The results showed that EforCSP3 was highly expressed in the female head,and its relative expression was much higher in adults than in other developmental stages.The fluorescence binding assays suggested that the EforCSP3 exhibited high binding affinities to a wide range of host-related volatiles,among which dibutyl phthalate,1-octene,β-elemene,and tridecane had the strongest binding affinity with EforCSP3,besides α-humulene and β-myrcene,and should be assessed as potential attractants.Protein structure modeling and molecular docking predicted the amino acid residues of EforCSP3possibly involved in volatile binding.α-Humulene and β-myrcene attracted E.formosa in a previous study and exhibited strong binding affinities with EforCSP3 in the current study.In conclusion,EforCSP3 may be involved in semiochemical reception by E.formosa.

Effects of ELF magnetic fields on protein expression profile of human breast cancer cell MCF7

Extremely Low Frequency Magnetic Fields (ELF MF) has been considered as a “possible human carcinogen” by International Agency for Research on Cancer (IARC) while credible mechanisms of its carcinogenicity remain unknown. In this study, a proteomics approach was employed to investigate the changes of protein expression profile induced by ELF MF in human breast cancer cell line MCF7, in order to determine ELF MF-responsive proteins. MCF7 cells were exposed to 50 Hz, 0.4 mT ELF MF for 24 h and the changes of protein profile were examined using two dimensional electrophoresis. Up to 6 spots have been statistically signifi-cantly altered (their expression levels were changed at least 5 fold up or down) compared with sham-exposed group. 19 ones were only detected in exposure group while 19 ones were missing. Three proteins were identified by LC-IT Tandem MS as RNA binding protein regulatory subunit、Proteasome subunit beta type 7 precursor and Translationally Controlled Tumor Protein. Our finding showed that 50 Hz, 0.4 mT ELF MF alternates the protein profile of MCF7 cell and may affect many physiological functions of normal cell and 2-DE coupled with MS is a promising ap-proach to elucidating cellular effects of electromagnetic fields.

Identification and expression profiles analysis of odorant-binding proteins in soybean aphid,Aphis glycines(Hemiptera:Aphididae)

The soybean aphid,Aphis glycines,is an extreme specialist and an important invasive pest that relies on olfaction for behaviors such as feeding,mating,and foraging.Odorant-binding proteins(OBPs)play a vital role in olfaction by binding to volatile compounds and by regulating insect sensing of the environment.In this work we used rapid amplification of complementary DNA ends technology to identify and characterize 10 genes encoding A.glycines OBPs(AglyOBPs)belonging to 3 subfamilies,including 4 classic OBPs,5 Plus-C OBPs,and one Minus-C OBP.Quantitative real-time polymerase chain reaction demonstrated variable specific expression patterns for the 10 genes based on developmental stage and aphid tssue sampled.Expression levels of 7 AglyOBPs(2,3,4,5,7,9,and 10)were highest in the 4th instar,indicating that the 4th nymphal instar is an important developmental period during which soybean aphids regulate feeding and search for host plants.Tissue-specific expression results demonstrated that AglyOBP2,7,and 9 exhibited significantly higher expression levels in antennae.Meanwhile,ligand-binding analysis of5 OBPs demonstrated binding of AglyOBP2 and AglyOBP3 to a broad spectrum of volatiles released by green leaf plants,with bias toward 6-to 8-carbon chain volatiles and strong binding of AglyOBP7 to trans-B-farnesene.Taken together,our findings build a foundation of knowledge for use in the study of molecular olfaction mechanisms and prov ide insights to guide future soybean aphid research.

Widely predicting specific protein functions based on protein-protein interaction data and gene expression profile

GESTs (gene expression similarity and taxonomy similarity), a gene functional prediction approach previously proposed by us, is based on gene expression similarity and concept similarity of functional classes defined in Gene Ontology (GO). In this paper, we extend this method to protein-protein interac-tion data by introducing several methods to filter the neighbors in protein interaction networks for a protein of unknown function(s). Unlike other conventional methods, the proposed approach automati-cally selects the most appropriate functional classes as specific as possible during the learning proc-ess, and calls on genes annotated to nearby classes to support the predictions to some small-sized specific classes in GO. Based on the yeast protein-protein interaction information from MIPS and a dataset of gene expression profiles, we assess the performances of our approach for predicting protein functions to “biology process” by three measures particularly designed for functional classes organ-ized in GO. Results show that our method is powerful for widely predicting gene functions with very specific functional terms. Based on the GO database published in December 2004, we predict some proteins whose functions were unknown at that time, and some of the predictions have been confirmed by the new SGD annotation data published in April, 2006.

油莎豆CePIP2;1的克隆、亚细胞定位与表达分析

油莎豆是一种在块茎中高水平积累油脂的草本油料作物,其未成熟块茎的含水量高达85%,水分平衡对于块茎的发育与代谢至关重要。质膜内在蛋白(PIP)是介导细胞间水分跨膜运输的主要通道。本研究报道1个块茎高水平表达的PIP基因CePIP2;1,该基因含有3个内含子,预测编码288个氨基酸(aa),其理论分子量(MW)为30.34 kDa,等电点(pI)为8.60,总平均疏水指数(GRAVY)为0.529,不稳定系数(Ⅱ)为29.60,属于典型的稳定、碱性、疏水型蛋白。CePIP2;1含有保守的MIP结构域,ar/R选择性滤器为F-H-T-R,Froger位点为Q-S-A-F-W,符合高水分转运活性PIP的特征。进化分析显示,CePIP2;1与OsPIP2;1、OsPIP2;2和OsPIP2;3聚在一起,序列相似性分别为91.72%、90.31%和84.98%,支持其归为PIP2亚类。亚细胞定位分析显示,CePIP2;1定位在烟草叶片的细胞膜。进一步的表达分析显示,虽然CePIP2;1为主要的块茎表达PIP基因,但其在叶片、叶鞘、匍匐茎和根等组织中的表达丰度更高,最高的为叶片,而在芽尖中的表达丰度与块茎相当;在块茎的4个典型发育时期(S_(1)~S_(4))中,CePIP2;1基因呈现先升后降的钟形趋势,表达丰度最高的是S_(2)。这些结果为解析油莎豆的水分平衡机制奠定基础。

油莎豆水通道蛋白基因CeTIP2;1的克隆与分析

油莎豆起源于非洲和地中海沿岸,是一种在块茎中高水平积累油脂的新型草本油料作物。水分平衡对于块茎的发育与代谢至关重要。液泡膜内在蛋白(TIP)是一类液泡膜定位并具有高效水分转运活性的水通道蛋白,包含TIP1~5等5个亚类。本研究基于油莎豆的基因组和转录组数据,采用RT-PCR技术对1个块茎高水平表达的TIP基因CeTIP2;1进行克隆。序列分析表明:CeTIP2;1的基因全长3323 bp,含有2个内含子,编码区长747 bp,编码248个氨基酸,理论分子量为24.73 kDa,等电点为5.09,总平均疏水指数为0.948,脂肪族指数为114.60,不稳定系数为21.76,属于稳定的酸性疏水型蛋白,与其液泡膜定位一致;该蛋白含有保守的MIP结构域,其中包括6个跨膜螺旋、2个半螺旋以及2个典型的NPA基序。CeTIP2;1与AtTIP2;1的序列相似性高达87.20%,远高于与SoPIP2;1的46.23%。进化分析进一步证实,CeTIP2;1隶属于TIP2亚类,是AtTIP2;1的直系同源基因。进化分析同时显示TIP2亚类早在单、双子叶植物分化之前就已经进化出2个小组。表达分析显示:CeTIP2;1在叶片、叶鞘、根、芽尖、匍匐茎、块茎等主要组织中均有较高水平的表达,丰度最高的是块茎和匍匐茎,最低的为芽尖;在块茎的发育过程中,呈现先升后降的钟形趋势,峰值出现在膨大中期,最低的为成熟期。此外,CeTIP2;1还在块茎的蛋白组中被检测到,表明其功能的重要性。这些结果为进一步揭示油莎豆的块茎水分平衡机制奠定坚实的基础。

春尺蠖热激蛋白AcinHsp70基因的克隆、分子特性与表达分析

【目的】克隆春尺蠖热激蛋白Hsp70基因,分析其理化性质与表达情况,探究该基因在春尺蠖应对外界不利环境中的作用。【方法】基于前期所测转录组数据(春尺蠖幼虫不同低温胁迫),克隆获取Hsp70基因,命名为AcinHsp70(GenBank登录号:OP750249);利用生物信息分析软件对Hsp70基因的理化性质进行分析;采用qPCR技术检测AcinHsp70基因在不同温度(-10、-5、0、5和25℃)胁迫中回温与不回温及4℃不同时间处理下的表达谱。【结果】春尺蠖AcinHsp70基因完整CDS序列全长1899 bp,编码633个氨基酸,蛋白质预测分子量为69.91 kDa,预测等电点为5.51。在N端均不含信号肽且无跨膜区。磷酸位点检测分别发现丝氨酸、苏氨酸和酪氨酸磷酸位点24、26和8个;未发现糖基化位点。该蛋白的平均亲水性为-0.507,预测不稳定系数为33.19,故该蛋白属于亲水稳定蛋白,亚细胞定位结果显示,其主要位于细胞质。序列一致性比对与系统进化树分析结果显示,春尺蠖与庆网蛱蝶(Melitaea cinxia)亲缘关系最近,序列一致性达92.91%。基因表达结果显示,不同温度(-10℃除外)胁迫下,AcinHsp70基因表达量均无明显差异,回温后表达量显著上调。AcinHsp70基因表达量总体呈先升后降趋势,在4℃处理1 h后表达量达到最大值。【结论】AcinHsp70基因与春尺蠖应对低温胁迫的响应密切相关,结果有助于揭示该基因在低温胁迫下的分子功能。

油莎豆块茎表达CeTIP4;1基因的克隆、亚细胞定位及表达分析

液泡膜内在蛋白(tonoplast intrinsic protein,TIP)是调控植物细胞内水分平衡的主要水通道蛋白类型,其根据进化关系可分为TIP1~TIP5等5个亚类。本研究基于前期获得的油莎豆全长转录组对1个块茎高水平表达的TIP基因(命名为CeTIP4;1)进行克隆和鉴定。序列分析显示,该基因含有2个保守的内含子,编码区全长753 bp,预测编码250个氨基酸,理论分子量为25.57 kDa、等电点(pI)为6.02、总平均疏水指数(GRAVY)为0.884,属于典型的酸性疏水型蛋白;该蛋白含有1个保守的MIP(major intrinsic protein)结构域,且具有已结晶AtTIP2;1类似的结构特征。进化分析显示,CeTIP4;1与AtTIP4;1、OsTIP4;1、OsTIP4;2和OsTIP4;3聚在一起,序列相似性为69.62%~82.94%,支持其划归为TIP4亚类。本研究在基因克隆的基础上进一步构建了其亚细胞定位载体pNC-Cam1304-CeTIP4;1。在烟草叶片中的瞬时过表达和共聚焦显微镜观察显示,CeTIP4;1定位在细胞膜。进一步的表达分析显示,CeTIP4;1具有明显的组织特异性,主要在块茎、根、匍匐茎和芽尖中表达;在起始期、膨大中期、膨大晚期和成熟期等4个典型的块茎发育时期中,CeTIP4;1基因的表达呈现先升后降的趋势,峰值出现在膨大中期,最低的是成熟期,与含水量趋势基本一致。本研究结果为深入解析油莎豆块茎的水分平衡机制奠定坚实的基础。

春尺蠖保幼激素结合蛋白AcinJHBP基因的克隆、分子特征与表达分析

[目的]探究AcinJHBP基因在春尺蠖生长发育与滞育中的作用。[方法]基于前期所测春尺蠖蛹转录组数据(非滞育蛹-滞育蛹与滞育蛹-滞育解除蛹),克隆获取JHBP基因,命名为AcinJHBP(GenBank登录号:OR860357);利用生物信息学软件分析AcinJHBP基因的序列特征及理化性质;采用qPCR技术检测AcinJHBP的时空表达(不同发育阶段和组织)情况。[结果]春尺蠖AcinJHBP基因完整CDS序列全长726 bp,编码241个氨基酸,蛋白质预测分子量为26.97 kDa,预测等电点为5.13。春尺蠖AcinJHBP基因预测分子式为C_(1211)H_(1915)N_(313)O_(364)S_(9),负电荷残基(Asp+Glu)总电荷为34,正电荷残基(Arg+Lys)总电荷为26,半衰期和脂肪指数分别为30 h和97.51。该蛋白的平均亲水性为-0.098,预测不稳定系数为27.13,故该蛋白属于亲水稳定蛋白,N端氨基酸为蛋氨酸(Met,M)。AcinJHBP在N端含一个长为17个氨基酸残基的信号肽,但无跨膜区;磷酸位点检测分别发现丝氨酸、苏氨酸和酪氨酸磷酸位点分别为9、6和3个,未发现糖基化位点。亚细胞定位结果显示,其主要位于细胞外。系统进化结果表明,AcinJHBP与同为鳞翅目尺蛾科的冬尺蠖(Operophtera brumata)血淋巴保幼激素结合蛋白JHBP亲缘关系最近。基因表达结果显示,在不同发育阶段,AcinJHBP基因在春尺蠖卵至5龄幼虫期间表达均较低(3龄幼虫除外),且无显著差异(P>0.05),而在非滞育蛹-成虫期,AcinJHBP表达量呈逐渐下降趋势。在不同组织中,春尺蠖AcinJHBP基因在雄虫头部表达量最高。[结论]AcinJHBP基因与春尺蠖生长发育与滞育密切相关,该结果为进一步揭示该基因在春尺蠖蛹越夏、越冬滞育中的功能奠定一定理论基础。

Cloning and Characterization of a Pathogenesis-Related Protein Gene TaPR10 from Wheat Induced by Stripe Rust Pathogen

Pathogenesis-related proteins （PRs） play many important roles in plant defense response against pathogen attack. To better understand the molecular mechanism of PR genes involved in wheat adult plant resistance （APR） to stripe rust, based on a differentially expressed transcribed derived fragment （TDF）, a novel PR gene from wheat cv. Xingzi 9104 infected by the Puccinia striiformis Westend f. sp. tritici Erikss. pathotype CY32, which was highly similar to the maize ZmPRIO gene and designated as TaPRIO, was identified using in silico cloning and RT-PCR method. This novel TaPRIO gene was predicted to encode a 160-amino acid protein with a deduced molecular weight of 17.06 kDa and an isoelectronic point （pI） of 5.19. An amino acid sequence analysis of TaPR10 demonstrated the presence of a typical conserved domain of pathogenesis related protein Bet v I family. Multiple alignment analysis based on the amino acids encoded by 10 different PRIO genes from maize （Zea mays）, rice （Oryza sativa）, broomcorn （Sorghum bicolor）, and wheat （Triticum aestivum） indicated that PR proteins of class 10 was conserved among the 4 plant species with about 80% similarity. DNA sequence of TaPRIO suggested the presence of one 84-bp intron with the splicing sites of GT-AT bi-nucleotide sequence between 188 and 271 bp. Using a real-time quantitative RT-PCR （qRT-PCR）, expression profiles of TaPRIO revealed that at the adult-plant stage, TaPRIO transcript was up-regulated as early as 12 h post-inoculation （hpi）, with the occurrence of maximum induction at 24 hpi. At the seedling stage, TaPRIO was also slightly induced 18 hpi. However, the transcript amount was relatively lower than that of the adult-plant stage. Taken together, these results suggest that TaPRIO may participate in wheat defense response of APR to stripe rust.

Identification and tissue distribution of odorant binding protein genes in Harmonia axyridis (Coleoptera:Coccinellidae)

The olfactory system of insects is crucial in modulating behaviors such as host seeking,mating,and oviposition.Odorantbinding proteins(OBPs)are involved in semiochemical recognition.OBPs recognize and bind odorants and transport them to odorant receptors located in olfactory neurons.Harmonia axyridis(Coleoptera:Coccinellidae)is a widely used predacious biological control agent for many agricultural and forestry pests.This study identified 19 OBPs in H.axyridis based on the antennal and whole-body transcriptomes of adults and obtained all the full-length open reading frames,including 11‘Classic’OBPs,7‘Minus-C’OBPs and 1‘Plus-C’OBP.They encoded 125 to 241 amino acid proteins with molecular weights ranging from 13.75 to 27.75 kDa and isoelectric points ranging from 4.15 to 8.80.Phylogenetic analyses were used to study the relationships between H.axyridis OBPs and OBPs from other species of Coleoptera.Quantitative real-time PCR(qPCR)analysis showed that HaxyOBP2,3,5,8,10,12,13,14,and 15 were highly expressed in antennae of both adult females and males.Moreover,HaxyOBP2,3,5,12,and 15 were more abundantly expressed in antennae than other body parts,while HaxyOBP13 and HaxyOBP14 were expressed predominantly,and at similar levels,in the head and antennae.The other OBP genes were highly expressed in non-olfactory tissues including the thorax,abdomen,legs,and wings.These results provide valuable information for further study of H.axyridis olfaction,which may ultimately enhance its use as a biocontrol agent.

Identification of differentially expressed genes in human uterine leiomyomas using differential display

In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down-regulated named M1 in leiomyoma) were confirmed by Northern analysis. Sequence comparison and Northern analysis proved that L1 is exactly the human ribosomal protein S19. It was present ubiquitously in 13 tissues tested but in various levels and even in different size. L1 was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPS19 overexpression might be a universal signal in rapid cell growth tissues.

Analysis of Genetic Variation of Seed Proteins in the Genus Vigna and among Its Relatives Cultivated in China

The genetic variation of seed proteins was assayed by SDSPAGE for 24 cultivars belonging to 5 species in Vigna and 7 species in its 7 relative genera cultivated in China. There were 48 polymorphic subunit bands discriminated from electrophoretic profiles. Two dendrograms were constructed by UPGMA cluster analyses using PHYLIP3.6 respectively. Variation among genera or species was larger than that among lower taxonomic categories level. Little variation among cuhivars of yardlong bean （Vigna sesquipedalis ） and small variation of lablab （ Lablab purpureus）, pea （Pisum sativum）, or sword bean （Canavalia gladiata）, but large variation of soybean or rice bean in their origin of China were all revealed. The seed proteins profiles of traditionally regarded as typical species in Vigna such as yardlong bean, rice bean and small bean were more similar than mungbean （Vigna radiata） and black gram （Vigna mungo） were. Mungbean and black gram had distinct seed proteins pattern, they should be of two species.

蛋白质芯片检测技术结合多维统计方法推断死亡时间

目的利用蛋白质芯片检测技术结合多维统计方法分析骨骼肌组织的蛋白质变化,进行死亡时间推断。方法大鼠颈椎脱臼处死后置于16℃环境中,提取死后不同时间点(0 d、1 d、2 d、3 d、4 d、5 d、6 d、7 d、8 d和9 d)骨骼肌的水溶性蛋白质,获取相对分子质量为14000~230000的蛋白质表达谱数据,采用主成分分析(principal component analysis,PCA)和正交偏最小二乘(orthogonal partial least squares,OPLS)进行数据分析,构建Fisher判别模型及反向传播(back propagation,BP)神经网络模型对大鼠死亡时间进行分类预测。另外收集不同死亡时间的人体骨骼肌蛋白质表达谱数据,通过热图和聚类分析观测其与死亡时间之间的关系。结果大鼠骨骼肌蛋白质谱峰随死亡时间呈一定的时序性变化。PCA结合OPLS判别分析结果显示,除死后6 d、7 d和8 d外,各死亡时间组间差异有统计学意义(P<0.05);经Fisher判别分析,模型内部交叉验证准确率为71.4%,外部验证准确率为66.7%;BP神经网络模型分类预测结果显示,模型内部交叉验证准确率为98.2%,外部验证准确率为95.8%。经聚类分析,人体骨骼肌蛋白质表达在死后4 d与死后25 h内差异有统计学意义。结论蛋白质芯片检测技术可快速、准确、高重复性地获取死后不同时间点大鼠和人体骨骼肌相对分子质量为14000~230000的水溶性蛋白质表达谱,建立基于多维统计方法的多种死亡时间推断模型,有望为死亡时间推断提供新的思路和方法。