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961.
Expression of a modified Cry1Ie gene in E. coli and in transgenic tobacco confers resistance to corn borer 总被引:1,自引:0,他引:1
Liu YJ Song FP He KL Yuan Y Zhang XX Gao P Wang JH Wang GY 《Acta biochimica et biophysica Sinica》2004,36(4):309-313
The wild-type Crylle gene from Bacillus thuringiensis was modified for its efficient expression in transgenic plants. Modified Crylle gene (designated as Cryllem) was cloned into prokaryotic expressionvector pET28b and its expression in E.coli was confirmed by SDS-PAGE analysis. Bioassays using crude expression products in E.coli revealed that CrylIem protein had a similar toxicity to corn borer as wild-type CrylIe. CrylIem gene was then inserted downstream of the maize ubiquitin-1 promoter in plant expression vector p3301. Transgenic tobacco plants carrying Cryllem showed insecticidal activity against corn borer. 相似文献
962.
Zhu H Zhou ZM Huo R Huang XY Lu L Lin M Wang LR Zhou YD Li JM Sha JH 《Acta biochimica et biophysica Sinica》2004,36(3):227-234
A gene, presumably involved in spermatogenesis, was identified and characterized by using cDNA microarray. Hybridization intensity was 2.13 fold higher in adult testis than that in fetal testis.The full length of this gene was 4288bp and it encoded a 578 amino acid protein. Conserved structure and amino acid sequence analysis revealed that the protein contained 1 Thif-domain, 2 UBACT-domains,and a functional active site cysteine lay upstream of UBACT domain, all of them also existed in ubiquitin-activating enzyme E1 and E1 like proteins. So we named this gene as a novel ubiquitin-activating enzyme E1 like gene (nUBE1L). Expression profile showed that nUBE1L was predominantly expressed in testis.Comparison of the expression of nUBE1L in different developmental stages of testis indicated that it was highly expressed in adult testis. In conclusion, nUBE1L is a novel human E1 like gene highly expressed inadult testis, which plays key role in ubiquitin system, and accordingly influences spermatogenesis and male fertility. 相似文献
963.
This paper examines recent developments and applications of Hidden Markov Models (HMMs) to various problems in computational biology, including multiple sequence alignment, homology detection, protein sequences classification, and genomic annotation. 相似文献
964.
The splicing of many alternative exons in the precursor messenger RNA (pre-mRNA) is regulated by extracellular factors but the underlying molecular bases remain unclear. Here we report the differential regulation of Bcl-x pre-mRNA splicing by extracellular factors and their distinct requirements for pre-mRNA elements. In K562 leukemia cells, treatment with interleukin-6 (IL-6) or granulocyte-macrophage colony stimulating factor (GM-CSF) reduced the proportion of the Bcl-xL variant mRNA while treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) had no effect. In U251 glioma cells, however, TPA efficiently increased the Bcl-xL level. These regulations were also seen for a transfected splicing reporter mini-gene. Further analyses of deletion mutants indicate that nucleotides 1-176 of the downstream intron are required for the IL-6 effect, whereas additional nucleotides 177-284 are essential for the GM-CSF effect. As for the TPA effect, only nucleotides 1-76 are required in the downstream intron. Thus, IL-6, GM-CSF and TPA differentially regulate Bcl-x splicing and require specific intronic pre-mRNA sequences for their respective effects. 相似文献
965.
Responses to oligogalacturonic acid (OGA) were determined in transgenic Arabidopsis thaliana seedlings express-ing the calcium reporter protein aequorin. OGA stimulated a rapid, substantial and transient increase in the concentration of cytosolic calcium ([Ca^2 ]cyt) that peaked after ca. 15 s. This increase was dose-dependent, saturating at ca. 50 μg Gal equiv/ml of OGA. OGA also stimulated a rapid generation of H202. A small, rapid increase in H2O2 content was followed by a much larger oxidative burst, with H2O2 content peaking after ca. 60 min and declining thereafter. Induction of the oxidative burst by OGA was also dose-dependent, with a maximum response again being achieved at ca. 50 μg Gal equiv/mL. Inhibitors of calcium fluxes inhibited both increases in [Ca^2 ]cyt and [H2O2], whereas inhibitors of NADPH oxidase blocked only the oxidative burst. OGA increased strongly the expression of the defence-related genes CHS,GST, PAL and PR-1. This induction was suppressed by inhibitors of calcium flux or NADPH oxidase, indicating that increases in both cytosolic calcium and H2O2 are required for OGA-induced gene expression. 相似文献
966.
967.
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. 相似文献
968.
多巴胺D1和D2受体激动剂和拮抗剂对脑缺血/再灌注损伤的影响 总被引:5,自引:0,他引:5
实验应用开阔法、组织病理学方法、原位末端标记(in situ terminal deoxynucleotidyl transferase-metliated de-oxy-UTP mick end labeling,TUNEL)法及免疫组织化学等方法,探讨多巴胺D1、D2受体激动剂和拮抗剂对沙土鼠前脑缺血/再灌注损伤海马CA1区神经元凋亡及凋亡相关基因bcl-2、bax表达的影响。结果显示:前脑缺血5min可引起沙土鼠探索活动增加;再灌注3d,海马CA1区约95%的锥体细胞凋亡;再灌注7d,海马CA1区仅残存约2%—7%的存活锥体细胞;前脑缺血5min可抑制bcl-2的表达并诱导bax表达增高;预先应用D2受体激动剂培高利特可减轻缺血后沙土鼠行为学异常、抑制海马CA1区锥体细胞凋亡、提高锥体细胞存活数、显著诱导bcl-2的表达并抑制bax的表达。预先应用SKF38393、SCH23390及螺哌隆对以上结果无明显影响。实验结果提示,培高利特具有确切的脑保护作用,诱导bcl-2并抑制bax的表达可能是其脑保护作用机制之一。 相似文献
969.
为研制肿瘤相关寡核苷酸芯片,并实现其在抗肿瘤反义核酸“癌泰得”作用机理研究方面的初步应用,制备了包含近450种肿瘤相关基因特异寡核苷酸探针的寡核苷酸芯片,建立了相应的质控标准.“癌泰得”用脂质体转染HepG2肿瘤细胞,提取细胞总RNA反转录并荧光标记cDNA,用制备的寡核苷酸芯片检测肝癌细胞HepG2的肿瘤相关基因表达水平,用软件分析获得其差异基因表达谱.0.4 μmol/L的反义核酸“癌泰得”作用于HepG2细胞15 h后,MDNCF、DHS等基因mRNA表达下调,MUC2、MPP11、LAT、HRIF-B、JNK3A1等mRNA基因表达上调,初步检测到了“癌泰得”的抗肿瘤作用可能的相关基因,为进一步的分子作用机理的探讨奠定基础.结果表明,制备的肿瘤相关芯片敏感度高、特异性高、重复性均较好,可用于检测肿瘤相关基因的表达谱,为临床诊断和基础研究提供了技术平台. 相似文献
970.
db/db小鼠糖尿病肾病相关基因的分析和克隆 总被引:5,自引:0,他引:5
用GM-U74A基因芯片分别检测了正常对照组(db/m小鼠)、糖尿病肾病组(db/db小鼠)、大黄酸治疗组(大黄酸150 mg/kg治疗12周)肾脏基因表达谱.发现在12 437个基因(包括表达序列标签)中,与正常对照组相比,糖尿病肾病组有1 085个基因表达下调,37个基因表达上调,其中变化幅度大于2倍,表达下调的有166个和表达上调的有29个.与糖尿病肾病组相比,大黄酸治疗组有384个基因表达下调,155个表达上调,其中变化幅度大于2倍,表达下调的有47个和表达上调的有30个.在此基础上,对其中的一个差异表达的表达序列标签(EST)进行了详细的生物信息学分析,发现它是一个未知功能基因——“REKEN cDNA 0610006H10”基因的一部分.在用RT-PCR进一步验证了其与糖尿病肾病的相关性后,对“REKEN cDNA 0610006H10”基因进行了克隆. 相似文献