长江华溪蟹精子超微结构的研究

透射电镜下观察长江华溪蟹成熟精子的形态和超微结构。精子呈椭球形,无鞭毛,具数条由核延伸而成的辐射臂精子。由球形的顶体,膜复合体,核杯及辐射臂组成,顶体由头帽,亚帽带,丁体囊及中央管组成。在中央管的基部发现有中心粒。核膜与质膜紧贴在一起形成了精子的界膜。     

小鼠γ干扰素/上皮生长因子受体结合域融合蛋白的构建及其抗肿瘤活性的提高

采用基因工程和蛋白质工程技术,构建了一种新颖的小鼠γ干扰素/上皮生长因子受体结合域融合蛋白,该蛋白保留了完全的抗病毒活性,达到10~8IU/L菌液;利用~(125)I-mEGF进行的受体结合竞争抑制实验,证明融合蛋白所带有的上皮生长因子结合域有与EGF竞争结合受体的功能;体外抗乳腺癌细胞增殖实验证明融合蛋白具有比干扰素母体分子更明显的抗肿瘤细胞增殖效应;在小鼠B16黑色素瘤模型上,融合蛋白和干扰素母体治疗后的2组小鼠,平均瘤体重量分别为1.6和2.6g,具有统计学差异.表明融合蛋白具有比干扰素母体分子更强的体内肿瘤生长抑制作用.     

叶绿素缺乏的大麦突变体PSⅡ光化学效率较高的原因(英)

在相同的叶绿素浓度下,叶绿素缺乏的大麦突变体的叶绿体在450～480nm和600～640nm波长范围的光吸收值比野生型略高,而在400～440nm和700～740nm波长范围的光吸收值明显高于野生型;突变体叶片和叶绿体的低温（77K）荧光发射强度较低,而两个光系低温荧光产量的比值（r685／F735）较高;失去Mg2 后,突变体和野生型的F685／F735和Fv／Fm均降低,但突变体的降低幅度较小;突变体的叶绿体中基粒片层数目较少、长度较短,而间质片层较长。这些结果表明,大麦突变体PSⅡ向PSⅠ的激发能转移较少是其PSⅡ光化学效率较高的重要原因。     

Expression of cytochrome P4502E1 in human liver: Relationship between genotype and phenotype in Chinese

Polymorphic cytochrome P4502E1 (CYP2E1) plays an important role in the metabolic activation of many carcinogens. We have previously shown that the c1/c1 genotype recognized byRsa I in the 5′-regulatory region of theCYP2E1 may be a susceptibility factor for developing esophageal cancer and lung cancer in Chinese. The present study was to investigate the relationship between theRsa I genotype and the expression of CYP2E1 in human livers. A total of 50 liver specimens were genotyped forCYP2E1 and assayed for CYP2E1 protein contents and functional activity by using specific antibody in immunoblot and a probe substrate,p-nitrophenol. A considerable interindividual variation in CYP2E1 protein (20-fold) and functional activity (56-fold) was observed among these liver samples. However, when they were categorized according to genotype, the mean content of CYP2E1 protein was significantly higher among individuals with the c1/c1 genotype than that among those having c1/c2 or c2/c2 genotype [124.0±83.9 pmol/mg (n = 28) versus 65.5 ±38.9 pmol/mg (n = 22),P<0.01]. The mean activity of CYP2E1 towardsp-nitrophenol for the c1/c1 genotype was also higher than that for the variant genotypes (198.4±27.8 pmol/min/mg versus 101.2 ±18.1 pmol^-1 · min^-1 · mg^-1,P<0.01). Also, the protein levels and functional activity showed a significant correlation (r = 0.68,P<0.01). These results demonstrate an association between theRsa I genotype and the phenotype of CYP2E1 in our samples, and the data are compatible with the assumption thatCYP2E1 c1/c1 genotype is a susceptibility factor for certain cancers in Chinese.     

STUDY ON THE GENETIC VARIATION OF THE COTTON BOLLWORM HELICOVERPA ARMIGERA (HÜBNER) POPULATIONS IN CHINA*

Abstract Population genetic variations of Helicoverpa armigera (Hiibner) over different major cotton growing regions were analyzed by DNA polymorphism amplified with four simple repetitive sequence primers. The results showed that the laboratory population had relatively lower genetic variation than natural populations. The genetic variation between natural populations was not significant and genetic variation existed in the same location from different years, indicating frequent migration among natural cotton bollworm populations. Cluster analysis indicated that individuals from Chaoyang of Liaoning Province (CY), Gaotang of Shandong Province (GT) and Dafeng of Jiangsu Province(DF) were more mixed each other, which suggested that CY population might have higher gene flow with GT and DF populations, especially with GT population. It supports the theory that the cotton bollworm in Northeast China came from Shandong and Hebei Provinces. This result also demonstrated that the molecular makers in this study are sensitive to detect population genetic structure changes.     

Cloning, sequencing and function ofsanA, a gene involved in nikkomycin biosynthesis ofStreptomyces ansochromogenes

Several genetically stable mutants blocked in nikkomycin biosynthesis were obtained after the slightly germinated spores ofStreptomyces ansochromogenes, a nikkomycin producer, were treated with ultra violet radiation. One of the mutants is the same in morpholotical differentiation as the wild type strain and is designated as NBB19. A DNA library was constructed using plasmid pIJ702 as cloning vector, NBB19 as cloning recipient. A 6 kb DNA fragment which can genetically complement NBB19 was cloned when screening the library for antifungal activity. Sequence analysis showed that the 3 kbBgl II -Sal I fragment contains one complete ORF (ORF1) and one partial ORF (ORF2). ORF1 is designated assanA. sanA is 1 365 bp, encoding a protein consisting of 454 amino acid residues. Database searching indicated thatsanA is homologous to the hypothetical methyltransferase inPyrococcus horikoshii with 25% identities and 41% positives. Disruptant ofsanA lost the ability to synthesize nikkomycin. It indicated thatsanA is a novel gene which is essential for nikkomycin biosynthesis.     

大肠杆菌CusS的生物信息学分析及对银离子胁迫的响应

【目的】解析大肠杆菌(Escherichia coli) K-12菌株同型二聚体内膜传感器组氨酸激酶(sensor histidine kinase, CusS)蛋白在细菌应答金属银离子胁迫中的调控机制,为该菌的防治提供重要科学依据。【方法】利用ProtParam、ProtScale、Protein-Sol、TMHMM、SignalP、LocTree3、NetNGlyc-1.0、NetPhosBac-3.0、SOPMA、I-TASSERF、STRING和MEGA分别预测CusS的理化性质、亲水性、可溶性、跨膜域、信号肽、亚细胞定位、糖基化位点、磷酸化位点、二级结构、三级结构、蛋白互作的关系网络和蛋白在革兰阴性杆菌中的同源性。采用Red同源重组技术构建大肠杆菌ΔcusS,在不同培养基中连续监测ΔcusS的生长情况,观察该基因缺失后的细菌生长活性;通过最小抑菌浓度(minimal inhibitory concentration, MIC)试验评价该缺失株对金属铜、银离子和临床常见抗生素的敏感性变化;运用RT-qPCR检测cusS缺失后其下游基因cusCFBA和cusR转录水平。【结果】CusS蛋白由480个氨基酸组成,相对分子质量为53 738.05,原子总数为7 624,等电点为6.02,具有稳定性,是一种亲水性、不溶性蛋白;含有跨膜域;不存在信号肽,定位于细胞内膜中;存在2个糖基化位点、24个丝氨酸磷酸化位点、14个苏氨酸磷酸化位点和3个酪氨酸磷酸化位点;二级结构中α-螺旋占比55.42%,β-折叠占比11.67%,β-转角占比3.75%,无规则卷曲占比29.17%;cusS在埃希菌属和志贺菌属中的保守性高;菌落PCR和一代测序验证ΔcusS构建成功;连续检测生长曲线表明cusS缺失并不影响细菌的生长代谢,但CusS蛋白为大肠杆菌抵御金属银胁迫的关键基因。【结论】cusS作为一个关键基因,它的缺失并不影响大肠杆菌的生长活性,但会显著降低细菌抵御银离子胁迫的应答能力。缺失cusS将使下游基因cusCFBA和cusR的mRNA表达水平显著下降。对CusS蛋白进行生物信息学分析及表型初探,为深入了解CusS在大肠杆菌应答银离子胁迫的调控机制奠定了基础。     

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Molecular Cloning of a Vernalization related cDNA Clone (Vrc) of Vrc79 in Winter Wheat (Triticum aestivum L.)

Vernalization is an essential factor in the flowering development of cold-required plants. There is a key stage of nucleic acid metabolism in the process of vernalization in winter wheat. To probe into the molecular determinants of vernalization, a cDNA library presumably enriching vernalization-related genes was prepared by specially expressed mRNAs from winter wheat ( Triticum aestivum L. cv. Jingdong No. 1) plumules at the key stage of 21 d vernalization being recovered as cDNAs after subtraction with mRNAs from nonvernalized and devernalized plumules. One vernalization-related cDNA clone (Vrc), Vrc79, which was only expressed at the key stage of 21 d vernalization, but not at other stages of nonvernalization, 4 d vernalization and devemalization, was isolated by differential screening of the library, and shown to be a vernalization-specific clone by Northern blot. Result of homology search suggested that Vrc79 was a novel gene identified in higher plant which was different from the cold-stress-induced genes and might play an important role in the floral induction in vernalization-requiring plants.