超滤法分级和纯化蛋白质的现状和前景(英文)

超滤法具有高产率、低成本、容易放大等显著优点,但其低选择性一直限制其在生物蛋白制品的分级和纯化中的应用。本文总结和分析了超滤法用于蛋白质分级和纯化的近期研究成果,对其低选择性的原因进行了分析,提出并论证了提高超滤选择性的技术途径。     

Molecular evidence for increased hematopoietic proliferation in the spleen of the b/b laboratory rat

The splenomegaly and the appearance of a significant number of CFU-E (erythroid colony-forming units) and BFU-E¹ (erythroid burst-forming units) in the Belgrade laboratory rat (b/b) spleen prompted us to analyse further the molecular evidence for increased hematopoietic proliferation in the b/b spleen. Messenger RNAs (mRNAs) specific for globins, proteins for iron transport and deposition and the band 3 protein were used in rat erythropoietic tissues as markers for proliferation and erythroid differentiation. In the b/b spleen, all mRNAs analysed display an erythroid-specific pattern of expression. This analysis also revealed an enhanced level of mRNA for ferritin in the +/b spleen, whereas erythrocyte-specific mRNA production was normal.     

Studies of a key protein in the mechanism of the excitation-contraction coupling process of frog skeletal muscle,using phenylglyoxal

The excitation-contraction (E-C) coupling process in single twitch fibres from frog toe muscle was inhibited selectively by phenylglyoxal (PGO), a specific guanidyl modifying reagent. A new protein (31.5 kDa), which has PGO-binding ability and seems to play a key role in the E-C coupling process, was solubilized from transverse tubule membrane-junctional sarcoplasmic reticulum complexes (TTM-JSR) of frog skeletal muscles, using¹⁴C-PGO. The monoclonal antibody against this protein applied extracellularly inhibited the E-C coupling process of the single fibres. This protein appears to constitute the very first step of input for E-C coupling. It is considered to behave as an indispensable part of an electrometer to measure membrane potentials. Therefore, the name electrometrin is suggested for the new protein.     

一个在梅山猪×大白猪杂交组合的背最长肌中差异表达的新基因的分离、cDNA 序列分析及组织表达谱

为了揭示猪杂种优势的分子机理,用mRNA差异显示技术从梅山猪×大白猪杂交组合的背最长肌中分离到一个差异表达的基因,并用半定量RT-PCR进行鉴定,随后通过cDNA末端快速扩增法(RACE)得到该基因的cDNA全长.经与GenBank进行Blast比较,这个基因与猪的已知的基因没有明显的同源性.基因预测显示这个基因编码一个由188个氨基酸组成的蛋白质,且该蛋白质具有保守的PRA1 family protein的结构域,同时该蛋白质和人、大鼠、小鼠的PRA1 family protein 3分别具有88%,88%,87%的同源性,因此将这个基因命名为猪的PRA1 family protein 3基因.进化树分析表明猪的PRA1family protein 3和人的PRA1family protein 3具有比大鼠、小鼠的PRA1family protein 3更近的亲缘关系.组织表达谱分析显示这个基因在肌肉与脂肪组织中表达丰富,在脾中中等表达,在心、肾、卵巢、肺中少量表达,在肝与小肠中几乎不表达.同时对这个基因的功能及与猪的杂种优势的关系进行了讨论.     

Inhibitory effect of the adenovirus type 5 E1A protein expressed in the yeast system

The human adenovirus type 5 E1A, a tumor- suppressor gene[1], codes for two major related proteins of 243 amino acids (12S) and 289 amino acids (13S) by al-ternative splicing in two exons[2]. Studies have been shown that E1A can regulate expression of many genes and cell cycle[3]. Both in vitro and in vivo experiments indicated that E1A could induce tumor cells differentia-tion, convert tumor cells into an epithelial phenotype, in-hibit tumor cell growth and metastasis and strongly en-ha…     

N-terminal of L protein of vesicular stomatitis virus contains a new signal sequence

The L protein (241kD) of vesicular stomatitis virus (VSV) is the most important snbnnit of the replication complex. The existence of specific localization signal in the L protein was investigated by making recombinant constructs expressing truncated mutants of the L protein fused to green fluorescent protein (GFP) in transient transfection assays. The chimeric genes encoding varied N-terminal of L and GFP gene were put under the control of T7 promoter or CMV promoter. The fusion proteins were transiently expressed in BHK-21, COS-7, CHO or Hep G2 cells. When more than 120 residues were deleted or only 96 residues were kept on the N-terminal, the fusion proteins were shown to be distributed throughout the cells, cytoplasm and nucleus under the confocal microscope. However, other chimeric proteins with 120 or more amino acids were dotted and distributed in the perinuclear regions. And the fusion protein with 96—120 aa has the similar distribution. A thirteen-residue peptide QGYSFLHEVDKEA (108—120) was identified as localization signal, whose function would be absolutely distributed with the deficiency of D or V. Our results show that there is an independent localizing signal in N-terminal domain of L protein of VSV and this functional signal is conserved in different cell lines.     

新疆北鲵肌肉蛋白质、氨基酸、无机元素及食物组成分析

本报道了新疆北鲵肌肉蛋白质含量为19．08％，氨基酸17种，无机元素有Ca等6种，成幼体食物组成中以石蚕及腺状介虫为主．     

酶法水解棉籽蛋白的制备与应用

以脱酚后的棉籽粉为原料 ,研究了酶解法制备可溶性棉籽水解蛋白的生产工艺。对其影响因素和产品的实际应用作了探讨。结果表明 ,利用中性蛋白酶AS1.398水解棉籽粉 ,产品得率可达 28.8%,蛋白质量分数为63.28%。棉籽水解蛋白可作为菌体发酵氮源。     

视黄醇结合蛋白基因的研究发展

视黄醇结合蛋白(Retinol-Binding Proteins，RBP)是一类维生素A的运载蛋白，参与血清和细胞内视黄醇／酸的转运，是疏水小分子结合蛋白家族的成员。RBP主要在肝脏中合成并释放入血液进而进入各种组织，通过与视黄醇、前白蛋白及细胞表面受体相互作用，在维生素A的储存、代谢、转运到周围靶器官中具有重要功能。细胞内视黄醇结合蛋白则主要在细胞内发挥类似作用。本文主要就血清及细胞内视黄醇结合蛋白的基因结构、作用机理、发育性表达、组织定位及染色体定位、对动物繁殖性能和胚胎发育的影响等方面进行综述。