Molecular biology international最新文献

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Role of B Cell Development Marker CD10 in Cancer Progression and Prognosis B细胞发育标志物CD10在肿瘤进展和预后中的作用

Molecular biology international

Pub Date : 2016-11-14 DOI: 10.1155/2016/4328697

D. Mishra, Sunita Singh, G. Narayan

The human CD10 antigen is a single pass, type II transmembrane, 100 kD cell surface glycoprotein belonging to peptidase M13 family. Identified in common acute lymphoblastic leukemia as a cancer specific antigen, CD10 is a cell surface ectoenzyme widely expressed on different types of cells. Earlier, it was used only as a cell surface marker to identify and differentiate between haematological malignancies. Later, reported to be present in various malignancies, it is thought to play significant role in cancer development and progression. Regulated expression of CD10 is necessary for angiogenesis and so forth. However its expression level is found to be deregulated in different cancers. In some cancers, it acts as tumor suppressor and inhibits tumor progression whereas in others it has tumor promoting tendency. However, its role in tumorigenesis remains unclear. This review summarises structural features, functions, and probable role of CD10 in cancer development.

人CD10抗原是一种单代、II型跨膜、100 kD的细胞表面糖蛋白，属于肽酶M13家族。CD10是一种广泛表达于不同类型细胞的细胞表面外酶，在常见的急性淋巴细胞白血病中作为癌症特异性抗原被发现。早些时候，它仅被用作细胞表面标记物来识别和区分血液系统恶性肿瘤。后来，它被报道存在于各种恶性肿瘤中，被认为在癌症的发生和进展中起重要作用。CD10的调控表达对于血管生成等是必需的。然而，它的表达水平在不同的癌症中被发现是不受控制的。在某些癌症中，它起到抑瘤作用，抑制肿瘤进展，而在另一些癌症中，它具有促瘤倾向。然而，它在肿瘤发生中的作用仍不清楚。本文综述了CD10的结构特征、功能及其在癌症发展中的可能作用。

引用次数: 41

Drosophila Enhancer of Rudimentary Homolog, ERH, Is a Binding Partner of RPS3, RPL19, and DDIT4, Suggesting a Mechanism for the Nuclear Localization of ERH 果蝇基本同源增强子ERH是RPS3、RPL19和DDIT4的结合伙伴，提示ERH核定位的一种机制

Molecular biology international

Pub Date : 2016-10-18 DOI: 10.1155/2016/8371819

S. Tsubota, Anthony C Phillips

The protein enhancer of rudimentary homolog, ERH, is a small, highly conserved protein that has been found in animals, plants, and protists. Genetic and biochemical interactions have implicated ERH in the regulation of pyrimidine biosynthesis, DNA replication, transcription, mRNA splicing, cellular proliferation, tumorigenesis, and the Notch signaling pathway. In vertebrates and insects, ERH is nuclearly localized; however, an examination of the ERH amino-acid sequence does not reveal any nuclear localization signals. In this paper we show that the first 24 amino acids contain sequences necessary and sufficient for nuclear localization. Through yeast two-hybrid screens, three new binding partners of ERH, RPS3, RPL19, and DDIT4, were identified. RPS3 was isolated from both human and Drosophila screens. These interactions suggest functions of ERH in cell growth, cancer, and DNA repair. The ERH sequences necessary for the interactions between ERH and RPS3 and RPL19 are mapped onto the same 24-amino-acid region in ERH which are necessary for nuclear localization, suggesting that ERH is localizing to the nucleus through binding to one of its DNA-binding partners, such as RPS3 or RPL19.

初级同源物的蛋白质增强子ERH是一种小的、高度保守的蛋白质，存在于动物、植物和原生生物中。遗传和生化相互作用表明ERH参与了嘧啶生物合成、DNA复制、转录、mRNA剪接、细胞增殖、肿瘤发生和Notch信号通路的调控。在脊椎动物和昆虫中，ERH是核定位的;然而，检查ERH氨基酸序列并没有发现任何核定位信号。在本文中，我们证明了前24个氨基酸包含核定位所必需和充分的序列。通过酵母双杂交筛选，鉴定出ERH的3个新的结合伙伴:RPS3、RPL19和DDIT4。从人类和果蝇的筛选中分离出RPS3。这些相互作用提示ERH在细胞生长、癌症和DNA修复中的功能。ERH与RPS3和RPL19相互作用所必需的ERH序列被映射到ERH中与核定位所必需的相同的24个氨基酸区域，这表明ERH是通过与dna结合伙伴之一(如RPS3或RPL19)结合而定位到细胞核的。

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引用次数: 10

Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4α Isoform in Humans TPM4α亚型在人体内的克隆、测序和表达

Molecular biology international

Pub Date : 2016-09-15 DOI: 10.1155/2016/3105478

D. Dube, S. Dube, L. Abbott, Ruham Alshiekh-Nasany, Charles Mitschow, B. Poiesz

In mammals, tropomyosin is encoded by four known TPM genes (TPM1, TPM2, TPM3, and TPM4) each of which can generate a number of TPM isoforms via alternative splicing and/or using alternate promoters. In humans, the sarcomeric isoform(s) of each of the TPM genes, except for the TPM4, have been known for a long time. Recently, on the basis of computational analyses of the human genome sequence, the predicted sequence of TPM4α has been posted in GenBank. We designed primer-pairs for RT-PCR and showed the expression of the transcripts of TPM4α and a novel isoform TPM4δ in human heart and skeletal muscle. qRT-PCR shows that the relative expression of TPM4α and TPM4δ is higher in human cardiac muscle. Western blot analyses using CH1 monoclonal antibodies show the absence of the expression of TPM4δ protein (~28 kDa) in human heart muscle. 2D western blot analyses with the same antibody show the expression of at least nine distinct tropomyosin molecules with a mass ~32 kD and above in adult heart. By Mass spectrometry, we determined the amino acid sequences of the extracted proteins from these spots. Spot “G” reveals the putative expression of TPM4α along with TPM1α protein in human adult heart.

在哺乳动物中，原肌球蛋白由四个已知的TPM基因(TPM1、TPM2、TPM3和TPM4)编码，每个基因都可以通过选择性剪接和/或使用替代启动子产生许多TPM亚型。在人类中，除了TPM4外，每个TPM基因的肌共聚异构体已经知道很长时间了。最近，在对人类基因组序列进行计算分析的基础上，预测的TPM4α序列已在GenBank中发布。我们设计了RT-PCR引物对，并在人类心脏和骨骼肌中表达了TPM4α转录本和一种新的TPM4δ亚型。qRT-PCR结果显示，TPM4α和TPM4δ在人心肌中相对表达量较高。使用CH1单克隆抗体进行Western blot分析显示，TPM4δ蛋白(~28 kDa)在人心肌中不表达。用相同抗体进行二维免疫印迹分析显示，在成人心脏中至少有9个分子量在32 kD及以上的不同原肌球蛋白分子的表达。通过质谱分析，我们确定了从这些斑点中提取的蛋白质的氨基酸序列。“G”点显示了成人心脏中TPM4α和TPM1α蛋白的推测表达。

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引用次数: 13

Molecular Characterization of Human Rotavirus from Children with Diarrhoeal Disease in Sokoto State, Nigeria 尼日利亚索科托州腹泻病儿童感染轮状病毒的分子特征

Molecular biology international

Pub Date : 2016-03-09 DOI: 10.1155/2016/1876065

B. Alkali, A. Daneji, A. A. Magaji, L. S. Bilbis, F. Bande

This study was conducted to detect and characterize prevalent human group A rotavirus strains from 200 diarrheic children in Sokoto, Nigeria, by ELISA, monoclonal antibody (Mab) serotyping and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) techniques. Rotavirus was detected in 25.5% of the children. The G-serotypes observed in circulation were G4: 16 (59.3%), G1: 4 (14.8%), G2: 3 (11.1%), G3: 3 (11.1%), and G12: 1 (3.7%). The monoclonal antibody (Mab) serotyping detected G1 and G3 but did not detect G4 and G2 serotypes. The Mab typing of the G1 and G3 serotypes was consistent with the result of the RT-PCR. The VP4 genotypes detected were P[6] 3 (13%), P[8] 11 (47.8%), and the rare human P genotype (P[9]), found in 9 patients (39.1%). Nine strains identified with the common G and P combinations were G4 P[8] 5 (56%), G4 P[6] 1 (11%), G1 P[8] 2 (22%), and G3 P[8] 1 (11%), while seven strains with unusual combinations or rare G or P genotypes identified were G12 P[8] 1 (14%), G2 P[8] 2 (29%), and G4 P[9] 4 (57%). To our knowledge this is the first molecular study of human rotavirus and report of rare human G and P serotypes in Sokoto State.

本研究采用ELISA、单克隆抗体(Mab)血清分型和逆转录聚合酶链反应(RT-PCR)技术对尼日利亚索科托200名腹泻儿童流行的人A组轮状病毒株进行检测和鉴定。25.5%的儿童检出轮状病毒。循环中g -血清型分别为G4: 16(59.3%)、G1: 4(14.8%)、G2: 3(11.1%)、G3: 3(11.1%)、G12: 1(3.7%)。单克隆抗体(Mab)血清分型检测G1和G3，但未检测G4和G2血清型。G1和G3血清型的单抗分型与RT-PCR结果一致。VP4基因型分别为p[6]3(13%)、p[9]1(47.8%)和罕见的人类P基因型(p[9])(39.1%)。G、P常见组合菌株为g4p[8](56%)、g4p[9](11%)、G1 P[8](22%)、G3 P[8]1 (11%)， G、P基因型异常组合菌株为g12p[8]1(14%)、G2 P[8]2(29%)、g4p[8](57%)。据我们所知，这是索科托州首次对人类轮状病毒进行分子研究，并报告了罕见的人类G和P血清型。

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引用次数: 10

Soluble Expression and Characterization of Biologically Active Bacillus anthracis Protective Antigen in Escherichia coli 生物活性炭疽芽孢杆菌保护性抗原在大肠杆菌中的可溶性表达及特性研究

Molecular biology international

Pub Date : 2016-02-07 DOI: 10.1155/2016/4732791

N. Suryanarayana, Vanlalhmuaka, Bharti Mankere, M. Verma, K. Thavachelvam, U. Tuteja

Bacillus anthracis secretory protein protective antigen (PA) is primary candidate for subunit vaccine against anthrax. Attempts to obtain large quantity of PA from Escherichia coli expression system often result in the formation of insoluble inclusion bodies. Therefore, it is always better to produce recombinant proteins in a soluble form. In the present study, we have obtained biologically active recombinant PA in small scale E. coli shake culture system using three different expression constructs. The PA gene was cloned in expression vectors bearing trc, T5, and T7 promoters and transformed into their respective E. coli hosts. The growth conditions were optimized to obtain maximum expression of PA in soluble form. The expression construct PA-pET32c in DE3-pLysS E. coli host resulted in a maximum production of soluble PA (15 mg L−1) compared to other combinations. Purified PA was subjected to trypsin digestion and binding assay with lethal factor to confirm the protein's functionality. Biological activity was confirmed by cytotoxicity assay on J774.1 cells. Balb/c mice were immunized with PA and the immunogenicity was tested by ELISA and toxin neutralization assay. This study highlights the expression of soluble and biologically active recombinant PA in larger quantity using simpler E. coli production platform.

炭疽芽孢杆菌分泌蛋白保护性抗原(PA)是炭疽亚单位疫苗的主要候选物。试图从大肠杆菌表达系统中获得大量PA，往往会导致形成不溶性包涵体。因此，以可溶性形式生产重组蛋白总是更好的。在本研究中，我们使用三种不同的表达结构在小型大肠杆菌摇培养系统中获得了具有生物活性的重组PA。将PA基因克隆到携带trc、T5和T7启动子的表达载体上，转化到各自的大肠杆菌宿主中。优化生长条件，以获得可溶性PA的最大表达量。与其他组合相比，在DE3-pLysS大肠杆菌宿主中表达PA- pet32c可溶PA产量最高(15 mg L−1)。纯化后的PA进行胰蛋白酶消化和致死因子结合试验，以确认蛋白的功能。对J774.1细胞进行细胞毒实验，证实生物活性。用PA免疫Balb/c小鼠，采用ELISA法和毒素中和法检测其免疫原性。本研究利用简单的大肠杆菌生产平台，大量表达了可溶性和生物活性的重组PA。

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引用次数: 5

SCAR Marker for Identification and Discrimination of Commiphora wightii and C. myrrha. SCAR 标记用于识别和区分 Commiphora wightii 和 C. myrrha。

Molecular biology international

Pub Date : 2016-01-01 Epub Date: 2016-03-16 DOI: 10.1155/2016/1482796

Pramod Kumar Sairkar, Anjana Sharma, N P Shukla

Commercially important Commiphora species are drought-tolerant plants and they are leafless for most of the year. Therefore, it is necessary to develop some molecular marker for the identification. Intended for that, in the present study, species-specific, sequence-characterized amplified regions (SCAR) markers were developed for proficient and precise identification of closely related species Commiphora wightii and C. myrrha, which may ensure the quality, safety, and efficacy of medicines made from these plants through adulterous mixing of these plants. Two species-specific RAPD amplicons were selected, gel-purified, cloned, and sequenced after screening of 20 RAPD primers. The sequence of 979 and 590 nucleotides (Genebank accession numbers K90051 and K90052) was used for development of 4 SCAR markers, namely, Sc1P, Sc1Pm, Sc2P, and Sc2Pm. Out of them, the Sc1Pm was specific for C. wightii, while Sc2P discriminated both the Commiphora species. These markers are first reported and will be useful for rapid identification of closely related Commiphora wightii and C. myrrha species.

具有重要商业价值的 Commiphora 品种是耐旱植物，一年中大部分时间都是无叶的。因此，有必要开发一些分子标记来进行鉴定。为此，本研究开发了具有物种特异性的序列特征化扩增区域（SCAR）标记，用于熟练、准确地鉴定近缘物种 Commiphora wightii 和 C. myrrha，从而确保这些植物制成的药物的质量、安全性和有效性，防止掺杂掺假。在对 20 个 RAPD 引物进行筛选后，选出了两个物种特异性 RAPD 扩增子，对其进行凝胶纯化、克隆和测序。979 和 590 个核苷酸的序列（基因库登录号 K90051 和 K90052）被用于开发 4 个 SCAR 标记，即 Sc1P、Sc1Pm、Sc2P 和 Sc2Pm。其中，Sc1Pm 对 C. wightii 具有特异性，而 Sc2P 则能区分这两个 Commiphora 种。这些标记为首次报道，将有助于快速鉴定近缘的 Commiphora wightii 和 C. myrrha 种。

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引用次数: 0

Total Protein Profile and Drug Resistance in Candida albicans Isolated from Clinical Samples. 临床分离的白色念珠菌的总蛋白谱和耐药性。

Molecular biology international

Pub Date : 2016-01-01 Epub Date: 2016-07-11 DOI: 10.1155/2016/4982131

Kamal Uddin Zaidi, Abin Mani, Vijay Thawani, Arti Mehra

This study was done to assess the antifungal susceptibility of clinical isolates of Candida albicans and to evaluate its total protein profile based on morphological difference on drug resistance. Hundred and twenty clinical isolates of C. albicans from various clinical specimens were tested for susceptibility against four antifungal agents, namely, fluconazole, itraconazole, amphotericin B, and ketoconazole. A significant increase of drug resistance in clinical isolates of C. albicans was observed. The study showed 50% fluconazole and itraconazole resistance at 32 μg mL(-1) with a MIC50 and MIC90 values at 34 and 47 and 36 and 49 μg mL(-1), respectively. All isolates were sensitive to amphotericin B and ketoconazole. The SDS-PAGE protein profile showed a prevalent band of ~52.5 kDa, indicating overexpression of gene in 72% strains with fluconazole resistance. Since the opportunistic infections of Candida spp. are increasing along with drug resistance, the total protein profile will help in understanding the evolutionary changes in drug resistance and also to characterize them.

本研究旨在评估临床分离的白色念珠菌的抗真菌敏感性，并根据其耐药性的形态差异评估其总蛋白谱。从不同临床标本中分离的120株白色念珠菌对氟康唑、伊曲康唑、两性霉素B和酮康唑4种抗真菌药物进行了药敏试验。临床分离的白色念珠菌的耐药性明显增加。结果表明，氟康唑和伊曲康唑对32 μg mL(-1)的耐药率为50%，MIC50和MIC90分别为34和47、36和49 μg mL(-1)。所有分离株均对两性霉素B和酮康唑敏感。SDS-PAGE蛋白谱显示~52.5 kDa的流行带，表明该基因在72%的氟康唑耐药菌株中过表达。由于念珠菌的机会性感染随着耐药性的增加而增加，总蛋白谱将有助于了解耐药性的进化变化并对其进行表征。

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引用次数: 17

Analyses of Physcomitrella patens Ankyrin Repeat Proteins by Computational Approach. 应用计算方法分析小壶菌锚蛋白重复序列。

Molecular biology international

Pub Date : 2016-01-01 Epub Date: 2016-06-27 DOI: 10.1155/2016/9156735

Niaz Mahmood, Nahid Tamanna

Ankyrin (ANK) repeat containing proteins are evolutionary conserved and have functions in crucial cellular processes like cell cycle regulation and signal transduction. In this study, through an entirely in silico approach using the first release of the moss genome annotation, we found that at least 54 ANK proteins are present in P. patens. Based on their differential domain composition, the identified ANK proteins were classified into nine subfamilies. Comparative analysis of the different subfamilies of ANK proteins revealed that P. patens contains almost all the known subgroups of ANK proteins found in the other angiosperm species except for the ones having the TPR domain. Phylogenetic analysis using full length protein sequences supported the subfamily classification where the members of the same subfamily almost always clustered together. Synonymous divergence (dS) and nonsynonymous divergence (dN) ratios showed positive selection for the ANK genes of P. patens which probably helped them to attain significant functional diversity during the course of evolution. Taken together, the data provided here can provide useful insights for future functional studies of the proteins from this superfamily as well as comparative studies of ANK proteins.

锚蛋白(ANK)重复序列是一种进化保守的蛋白，在细胞周期调控和信号转导等关键细胞过程中具有重要作用。在这项研究中，利用首次发布的苔藓基因组注释，我们发现至少54种ANK蛋白存在于P. patens中。基于不同结构域的组成，鉴定的ANK蛋白被划分为9个亚家族。对不同ANK蛋白亚家族的比较分析表明，除了具有TPR结构域的ANK蛋白亚群外，patens几乎包含其他被子植物物种中发现的所有已知ANK蛋白亚群。使用全长蛋白序列的系统发育分析支持亚家族分类，其中同一亚家族的成员几乎总是聚集在一起。同义分化比(dS)和非同义分化比(dN)均表现为ANK基因的正向选择，这可能有助于它们在进化过程中获得显著的功能多样性。综上所述，本文提供的数据可以为该超家族蛋白的未来功能研究以及ANK蛋白的比较研究提供有用的见解。

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引用次数: 2

FOXO3a Gene Polymorphism Associated with Asthma in Indian Population 印度人群中FOXO3a基因多态性与哮喘相关

Molecular biology international

Pub Date : 2015-12-09 DOI: 10.1155/2015/638515

S. Barkund, Tejas Shah, Nikhil Ambatkar, M. Gadgil, K. Joshi

Asthma is a chronic inflammatory disorder delineated by a heightened immunological response due to environmental or genetic factors. Single nucleotide polymorphism studies have shown that FOXO3a plays a pivotal role in maintaining immunoregulation. Polymorphism in FOXO3a has been linked to inflammatory diseases such as chronic obstructive pulmonary disease (COPD), Rheumatoid Arthritis, and Crohn's disease suggesting that FOXO3a may be associated with asthma. Airway inflammation in asthma is characterized by activation of T helper type 2 (Th2) T cells and Foxo family members are reported to play critical roles in the suppression of T cell activation. Thus this study was undertaken to investigate an association between single nucleotide polymorphism of the FOXO3a (rs13217795, C>T transition) gene and asthma in Indian population. To our knowledge we are the first ones reporting an association between FOXO3a and asthma.

哮喘是一种慢性炎症性疾病，由环境或遗传因素引起的免疫反应增强所描述。单核苷酸多态性研究表明，FOXO3a在维持免疫调节中起着关键作用。FOXO3a多态性与慢性阻塞性肺疾病(COPD)、类风湿性关节炎和克罗恩病等炎症性疾病有关，这表明FOXO3a可能与哮喘有关。哮喘气道炎症的特征是辅助性T型2 (Th2) T细胞的激活，据报道Foxo家族成员在抑制T细胞激活中起关键作用。因此，本研究旨在探讨印度人群中FOXO3a (rs13217795, C>T过渡)基因单核苷酸多态性与哮喘的关系。据我们所知，我们是第一个报道FOXO3a和哮喘之间关联的人。

引用次数: 22

Evaluation of Loop-Mediated Isothermal Amplification Assay for the Detection of Pneumocystis jirovecii in Immunocompromised Patients 环介导等温扩增法检测免疫功能低下患者肺囊虫的评价

Molecular biology international

Pub Date : 2015-11-19 DOI: 10.1155/2015/819091

Preeti Singh, Sundeep Singh, B. Mirdha, R. Guleria, S. Agarwal, A. Mohan

Pneumocystis pneumonia (PCP) is one of the common opportunistic infection among HIV and non-HIV immunocompromised patients. The lack of a rapid and specific diagnostic test necessitates a more reliable laboratory diagnostic test for PCP. In the present study, the loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of Pneumocystis jirovecii. 185 clinical respiratory samples, including both BALF and IS, were subjected to GMS staining, nested PCR, and LAMP assay. Of 185 respiratory samples, 12/185 (6.5%), 41/185 (22.2%), and 49/185 (26.5%) samples were positive by GMS staining, nested PCR, and LAMP assay, respectively. As compared to nested PCR, additional 8 samples were positive by LAMP assay and found to be statistically significant (p < 0.05) with the detection limit of 1 pg. Thus, the LAMP assay may serve as a better diagnostic tool for the detection of P. jirovecii with high sensitivity and specificity, less turn-around time, operational simplicity, single-step amplification, and immediate visual detection.

肺囊虫性肺炎(PCP)是HIV和非HIV免疫功能低下患者常见的机会性感染之一。由于缺乏快速和特异性的诊断测试，因此需要更可靠的PCP实验室诊断测试。本研究评价了环介导等温扩增(LAMP)法检测乙氏肺囊虫的效果。185份临床呼吸样本，包括BALF和IS，进行GMS染色、巢式PCR和LAMP测定。185份呼吸道样本中，GMS染色阳性12/185(6.5%)，巢式PCR阳性41/185 (22.2%)，LAMP法阳性49/185(26.5%)。与巢式PCR相比，LAMP法阳性的样品增加了8份，差异有统计学意义(p < 0.05)，检出限为1 pg。因此，LAMP法具有灵敏度高、特异性强、处理时间短、操作简便、一步扩增、直观检测等优点，可作为一种较好的诊断工具。

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引用次数: 13

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