土耳其斯坦东毕吸虫磷酸丙糖异构酶基因的克隆及其序列分析

目的 克隆土耳其斯坦东毕吸虫磷酸丙糖异构酶的全长基因。方法 根据日本血吸虫和曼氏血吸虫磷酸丙糖异构酶基因的保守区设计引物，利用RT—PCR扩增出土耳其斯坦东毕吸虫磷酸丙糖异构酶基闪的大片段，再结合RACE技术分别得到磷酸丙糖异构酶基因的3’端和5’端，将3部分序列拼接后获得磷酸丙糖异构酶基因全长cDNA序列，并提交GenBank。结果 成功克隆了土耳其斯坦东毕吸虫磷酸丙糖异构酶基因全长cDNA序列并提交GenBank，登录号为DQ092331。结论 土耳其斯坦东毕吸虫磷酸丙糖异构酶基因全长cDNA的克隆为进一步表达及其生物学性能的分析提供了理论基础。     

Folding and function of the myelin proteins from primary sequence data.

To explain how the myelin proteins are involved in the organization and function of the myelin sheath requires knowing their molecular structures. Except for P2 basic protein of PNS myelin, however, their structures are not yet known. As an aid to predicting their molecular folding and possible functions, we have developed a FORTRAN program to analyze the primary sequence data for proteins, and have applied this to the myelin proteins in particular. In this program, propensities for the secondary structure conformations as well as physical-chemical parameters are assigned to the amino acids and the pattern of these parameters is examined by calculating their average values, autocorrelation functions and Fourier transforms. To compare two proteins, their sequences are aligned using a unitary scoring matrix, and homologies are searched by plotting a two-dimensional map of the correlation coefficients. Comparison of the corresponding myelin basic proteins (MBP) and P0 glycoproteins (P0) for rodent and shark showed that the conserved residues included most of the amino acids which were predicted to form the alpha or beta conformations, while the altered residues were mainly in the hydrophilic and turn or coil regions. In both rodent and shark the putative extracellular domain of P0 glycoprotein displayed consecutive peaks of beta propensity similar to that for the immunoglobulins, while the cytoplasmic domain showed alpha-beta-alpha folding. To trace the immunoglobulin fold along the P0 sequence, we compared the beta propensity curve of P0 with that of the immunoglobulin M603, whose three-dimensional structure has been determined. We propose that the flat beta-sheets of P0 are orientated parallel to the membrane surface to facilitate their homotypic interaction in the extracellular space. An extra beta-fold in the extracellular domain of shark P0 compared with rodent P0 was found, and this may result in a greater attraction between the apposed extracellular surfaces and may account for a smaller extracellular space as measured by x-ray diffraction. A computer search of the myelin protein sequences for functional motifs revealed sites for N-glycosylation, phosphorylation, nucleotide binding, and certain enzyme activities. We note especially that there are potential nucleotide binding sites in proteolipid protein (PLP), MBP and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP). This is consistent with the experimental observations that PLP acts like an ionophore or proton channel when reconstituted into planar lipid bilayers, MBP binds GTP, and CNP catalyzes in vitro the hydrolysis of 2',3'-nucleotides into corresponding 2'-nucleotides.     

HLA-DPB1 gene polymorphism and multiple sclerosis: a large case-control study in the southwest of France

The polymorphism at the HLA-DPB1 locus has been characterized in a large number of patients with multiple sclerosis (n = 112) and in healthy controls (n = 115). Both patients and controls lived in the southwest of France (in the Pyrénées Atlantiques) and had similar ethnic background. The typing procedure involved the selective amplification of the second exon of the DPB1 locus by polymerase chain reaction, followed by hybridization of the amplified DNA with 14 sequence-specific oligonucleotide probes. Individual alleles were identified by the pattern of hybridization of the different probes. The distribution of the DPB1 alleles was not significantly different in multiple sclerosis patients and controls (p = 0.11). This does not corroborate the reported association of multiple sclerosis with the primed lymphocyte typing (PLT)-defined DPw4 specificity and is not in favour of a role played by polymorphic residues of the DP molecule in susceptibility to multiple sclerosis.     

New scoring matrix for amino acid residue exchanges based on residue characteristic physical parameters

Based on residue characteristic physical parameters, a new scoring matrix, called EMPAR, for amino acid exchanges in proteins was obtained. When comparing protein sequences for detecting homologies, the use of this matrix in place of the Dayhoff log-odds matrix yields results that reflect the topological similarities in the proteins. The use of EMPAR is equivalent to the parametric correlation coefficient approach of Ooi and his colleagues. This matrix correlates at 0.63 with the Dayhoff matrix.     

颈性眩晕的分型治疗及疗效分析

目的 将颈性眩晕分为上、下颈性眩晕，采取不同治疗方法并与常规治疗方法的疗效比较，以寻求更好的治疗方法。方法 将135例颈性眩晕随机分为试验组和对照组，对其治愈率及治疗时间进行统计学分析。结果 试验组的治愈率高于对照组，而试验组的治疗时间短于对照组，均有统计学意义。结论 对颈性眩晕进行分型并采用不同的治疗方法能提高治愈率，缩短治疗时间。     

一种识别蛋白质的保守氨基酸残基的新方法

保守氨基酸残基在生物进化过程中具有较强的稳定性,一般不会发生太大的变化;生物信息学认为：氨基酸序列决定蛋白质结构,蛋白质结构决定蛋白质功能。因此,作者基于氨基酸的生化特性、几何特性和动态特性描述了一个新颖的算法去发现蛋白酶家族中的保守氨基酸残基,而保守氨基酸残基将对蛋白质的结构和功能的研究具有重要的意义。     

C端序列的删除对非出血重组蛇毒纤溶酶的影响

[目的]研究C端序列对非出血重组纤溶酶(rFⅡ)特异性、活性和热敏感性的影响.[方法]通过SOEing PCR方法构建删除C端序列的突变体,突变体和rFⅡ分别在P.pastoris中诱导表达.表达和纯化的蛋白用SDS-PAGE和Westeren blot进行鉴定.之后分析其生化特性.[结果]rFⅡ及突变体通过SDS-PAGE和Westeren blot得到证实.生化分析揭示:①突变体对显色底物N-(p-Tosyl)-Gly-Pro-Lys-pNA的催化效率(Kcat/Km)是rFⅡ的1.9倍;②突变体和rFⅡ对氧化的胰岛素B链拥有共同的优先裂解位点,可是在随后的裂解中开始展现差异;③突变体显示了更高的纤(原)活性;④热处理表明突变体相较r FⅡ对温度的增加更敏感;⑤通过圆二色谱测量,突变体的螺旋比例有所增加.[结论]删除的序列参与rFⅡ的特异性、活性和热敏感性,该序列可作为下一步优化的候选序列.     

A study of the value of electrophoretic and other techniques for typing Acinetobacter calcoaceticus

Forty-four isolates of Acinetobacter calcoaceticus var anitratus collected during hospital outbreaks were studied using polyacrylamide gel electrophoresis (PAGE), plasmid analysis, antibiograms and biochemical tests to determine their degree of similarity. Reproducibility tests were also carried out on the PAGE and biochemical techniques to determine their validity when used to compare bacteria of the same type isolated intermittently. PAGE data was analysed densitometrically and isolates compared using a similarity matrix. All methods were able to subdivide the isolates, but results did not always correlate well between methods. Reproducibility data indicated that careful attention to technique is required when organisms are examined by PAGE sequentially. Results suggest that no single biotyping technique is likely to be adequate and that electrophoretic, biochemical and antibiogram data may complement one another and other epidemiological data in the typing of these organisms.