真核细胞的基因的组织

一、真核细胞基因的基本结构 1.转录单位: 从已知的数十种基因的顺序,可得出一个具有功能的基因的共同规律,在基因5’端-25至-75区,有CCAAT和TATAAA区(后者又称ATA box或Hogness box),相当于促进子区(Promotor),为体外转录所必需。     

没有神经的腿的再生

对于嵘螈肢体的再生,神经系统特别是切割后的残肢裹的神经纤维,一向是被认为有决定性意义的。晚近利用成体或接近变态的幼体的研究证明了:在再生芽基形成之后,切断神经,已经形成的芽基仍然可以继续发育成正常的肢体;但在再生芽基形成之     

审美的眼 怜悯的心

面向公众宣传教育需要考虑受众的兴趣。大熊猫的保护,和它呈现的很可爱的形象关系很大．人们不由得产生一种怜惜的情感．想要去保护它。反观扬子鳄．如果对它难以拍出那样可爱的图片．写不出那样煽情的文章．那就没什么影响。毕竟,人们怜惜可爱的动物．而对丑陋动物的境遇不太关心,这可能是我们人类演化产生和积淀下来的一种情感反应。     

知识的载体 智慧的力量

2009年5月,繁花烂漫的季节,高等教育出版社迎来了55周年华诞,《生命世界》杂志也走过了5年不平凡的历程。我们出版的高质量的教材和学术著作为我国的生命科学人才培养和学科建设发挥了积极的重要作用,受到了广大师生和科研人员的欢迎。《生命世界》在传播生命科学知识、提高大众科学素养方面起到了很好的作用,全国人大常委会副委员长、中国科学院院长路甬祥院士和全国人大常委会副委员长、中国科协主席、     

知识的载体 智慧的力量

2009年5月,繁花烂漫的季节,高等教育出版社迎来了55周年华诞,《生命世界》杂志也走过了5年不平凡的历程。我们出版的高质量的教材和学术著作为我国的生命科学人才培养和学科建设发挥了积极的重要作用,受到了广大师生和科研人员的欢迎。《生命世界》在传播生命科学知识、提高大众科学素养方面起到了很好的作用,全国人大常委会副委员长、中国科学院院长路甬祥院士和全国人大常委会副委员长、中国科协主席、     

真正的科学家真正的爱国者

卢永根院士是著名的作物遗传学家。他致力于水稻遗传育种研究,取得了显著的成绩。在青年时代,他以极大的热情投身到革命洪流中,既做地下党的工作,又刻苦地在岭大深造;在岭大毕业后至今50余年中,他在高校既教书,又从事科学研究,为人才培养和水稻遗传育种工作做出贡献。他总结的五点体会和崇高的思想境界:“我的青春年华已经献给党的科学事业,我准备把晚年继续献给这个事业。”令人深受启发,值得学习。经作者同意,现将刊登在中国科学院院士工作局《学部通讯》2003年第4期上的“院士自述:我的成长经历”这篇报道,转载于后。     

干酪生产的凝乳酶的开发

用于干酪制造的重组凝乳酶的开发处于激烈竞争中。美国Genencor公司和Chr.汉逊实验公司协作,在重组凝乳酶的商品化工作中取得显著进展。美国Collaborative研究公司(CR)用重组生物制造凝乳酶方法已取得美国专利局许可的专利权。     

人类胚胎的免疫系统与成人的有着截然不同的起源

UCSF的研究人员首次向我们证实,人类胚胎的免疫系统与成人的免疫系统有着截然不同的起源.相对于成人的免疫系统而言,胚胎免疫系统对待其环境中的外源物质更倾向于适应,而不是去消灭它.这一发现不但使我们能够更加了解新生儿是如何对感染以及疫苗反应的,而且可以用来解释许多难题,比如为何HIV     

方兴未艾的古代DNA的研究

保留在古代生物遗骸中的遗传物质DNA是一种重要的遗传资源。古代DNA的研究对于了解包括人类在内的各种生物的起源、进化和迁徙有重要意义。古代DNA的研究有其自身的特点,并且已经取得一系列重要成就。本文综述古代DNA研究的历史、方法和进展。 Abstract：DNA present in ancient samples can be recovered,amplified and analysed.It opens a new window for genetic analysis in many different disciplines,such as anthropology,archaeology,human population genetics,animal and plant evolutionary taxonomy and forensic science.In general,ancient DNA is rare in quantity,damaged in quality.To ensure the reproducibility and reliability of the results,great cares should be taken,such as various measurements against contamination and phylogenetic analysis of ancient DNA sequences.In this paper we review recovery,amplification and analysis of ancient DNA,also discuss the guidelines to ensure the authenticity of ancient DNA and the recent advances in ancient DNA study.     

有前途的基因工程的飞跃发展

试图在分子水平上解释生命现象的分子生物学从其工程学角度派生出了生物工程学,1972年,DNA重组技术取得了引人注目的成功。自那以后,这二方面就开始了惊人的发展,在这里,让我们来回顾一下过去的业迹。     

香石竹花瓣中乙烯诱导合成的60kD蛋白功能的初步研究

越来越多的研究表明植物器官的衰老与蛋白质合成密切相关[1～3]。乙烯在许多花卉衰老和一些水果的成熟过程中起重要作用[1,3],实验结果显示花对乙烯的反应依赖于新的蛋白质合成[2～5]。深入研究那些与衰老相关的蛋白质,特别是乙烯诱导合成的蛋白质的生化功能,对于了解掌握...     

盐胁迫下苜蓿中盐蛋白的诱导产生

盐胁迫下苜蓿叶片中蛋白质的合成受到抑制,而其离体叶绿体中蛋白质合成增强,ABA阻碍了后者的蛋白质合成。NaCl胁迫下,“松江”和“肇东”两品种的根和叶中均无新多肽出现。在盐敏感的“松江”品种离体叶绿体中,NaGl诱导70,65,60和43kD4种多肽产生,ABA诱导60和17kD两种多肽产生;在较抗盐的“肇东”品种离体叶绿体中,NaGl诱导83,80kD和43kD3种多肽产生,但100mmol/L NaCl并不诱导83kD多肽出现,ABA无明显作用。两品种的43kD多肽和肇东品种的80kD多肽都存在于类囊体膜上,而松江品种的60kD多肽则存在于叶绿体间质中。     

大肠杆菌脑微血管内皮细胞侵袭基因ibeB的编码产物为外膜蛋白前体

为进一步探讨大肠杆菌脑微血管内皮细胞侵袭基因ibeB的生物学特性 ,将ibeB基因克隆到pET2 8a(+)载体 ,以E .coliBL2 1 (DE3)为宿主菌 ,经IPTG诱导后 ,通过Ni2 + NTA树脂提纯IbeB蛋白 .SDS PAGE确定纯化蛋白的分子量 ;应用无蛋白酶的体外转录和翻译系统进一步鉴定ibeB基因表达蛋白的分子量 ;通过 [3 5S]Met标记的体内T7表达体系并结合膜蛋白分离技术定位IbeB蛋白在细菌中的亚细胞分布 ;利用细菌侵袭实验分析IbeB蛋白抗体对E .coliK1侵袭人脑微血管内皮细胞的封闭作用 .结果发现 ,ibeB基因的重组蛋白表达纯化产物呈现出 5 0kD和 34kD两种分子量大小 ,5 0kD存在于表达细菌的可溶性部分 ,而 34kD则存在于包涵体中 ;体外翻译实验也显示出较弱的 5 0kD和较浓的 34kD两个蛋白带 ;体内T7表达体系实验显示 34kD的IbeB成熟蛋白定位于E .coli的外膜 ;抗 34kDIbeB蛋白抗体能封闭E .coli对人脑微血管内皮细胞的侵袭 .这些结果提示 ,大肠杆菌脑微血管内皮细胞侵袭基因ibeB的编码产物为 5 0kD的外膜蛋白前体 ,该前体可通过分子内剪接形成成熟的 34kDIbeB蛋白     

Studies on the physiological function of spermine in the process of progesterone induced toad oocyte maturation

Spermidine or spermine but not putrescine inhibited progesterone induced Bufo bufo gargarizans oocyte maturation.The ID50 for spermine inhibition via intra -oocyte microinjection on maturation induced by progesterone was 6.8mM(100nl).Spermine could inhibit MPF induced toad oocyte maturation with a much higher ID50.A 55 kD protein was dephosphorylated during the process of progesterone induced oocyte maturation .Spermine selectively promoted the level of phosphorylation of this protein in both progesterone-stimulated and hormone-untreated oocytes.The extent of its dephosphorylation was fairly Correlated with the percentage of GVBD in the hormone stimulated oocytes.The level of endogenous spermine was reduced by 28% between the perod of 0.40 GVBD50 and 0.60 GVBD50,at which 55 kD protein was dephosphorylated.Spermine inhibited progesterone-stimulated protein synthesis in almost the same dose dependent manner as its inhititory effect on the hormone-induced maturation,The endogenous spermine regulated 55 kD protein dephosphorylation which may trigger the increase of protein dephosphorylation which may trigger the increase of protein synthesis and in turn promote the activation of MPF,It is possible that 55 kD protein may be one of the components of messenger ribonucleoprotein(mRNP) particles.     

Identification of a basic glycoprotein induced by ethylene in primary leaves of azuki bean as a cationic peroxidase.

Ethylene causes the accumulation of seven different proteins (each designated AZxx according to its molecular mass, xx in kD) in excised primary leaves of azuki bean (Vigna angularis) (F. Ishige, H. Mori, K. Yamazaki, H. Imaseki [1991] Plant Cell Physiol 32: 681-690). A complementary DNA encoding an ethylene-induced basic glycoprotein, AZ42, from azuki bean was cloned and its complete nucleotide sequence was determined. Characterization of the cDNA was accomplished by monitoring expression of an immunoreactive protein in Escherichia coli that harbored the cDNA and by the identification of a partial amino acid sequence that was the same as that determined from the purified protein. An open reading frame (1071 base pairs) in the cDNA encoded a protein of 357 amino acids with a molecular mass of 39.3 kD. The amino acid sequence contained three regions that are highly conserved among peroxidases from eight different plants. Purified AZ42 exhibited peroxidase activity. The basic glycoprotein induced by ethylene was identified as a cationic isozyme of peroxidase. The corresponding mRNA was not present in leaves that had not been treated with ethylene, but it appeared after 1 h of treatment with ethylene and its level increased for the next 15 h. Accumulation of the mRNA was also induced after wounding or treatment with salicylate. The wound-induced increase in the level of the mRNA was suppressed by 2,5-norbornadiene, but the salicylate-induced increase was not.     

解毒酶基因cDNA克隆和高效表达

昆虫抗药性的一个重要机制是其产生的解毒酶可以将大剂量的农药脱去毒性[1] 。酯酶活性升高是库蚊对有机磷杀虫剂抗性的主要机制 ,与酯酶Ｂ1有关的抗性最高[2 ,3] 。酯酶与有机磷杀虫剂有非常强的结合力 ,可以迅速与之形成强结合体[4 ] 。酯酶的解毒作用具有很高的手性专一性 ,在有机磷化合物 ,特别是高毒的有机磷化合物的解毒作用中非常重要[1] 。高效表达解毒酶基因 ,将昆虫解毒酶用于人畜解毒的目的研究还未见报道。本文报道在大肠杆菌中高效表达昆虫解毒酶 ,并将产物用于实验动物有机磷中毒的解毒研究 ,为昆虫抗性相关基因的开发利用提…     

The human U4/U6 snRNP contains 60 and 90kD proteins that are structurally homologous to the yeast splicing factors Prp4p and Prp3p.

Immunoaffinity-purified human 25S [U4/U6.U5] tri-snRNPs harbor a set of polypeptides, termed the tri-snRNP proteins, that are not present in Mono Q-purified 20S U5 snRNPs or 10S U4/U6 snRNPs and that are important for tri-snRNP complex formation (Behrens SE, Lührmann R, 1991, Genes & Dev 5:1439-1452). Biochemical and immunological characterization of HeLa [U4/U6.U5] tri-snRNPs led to the identification of two novel proteins with molecular weights of 61 and 63kD that are distinct from the previously described 15.5, 20, 27, 60, and 90kD tri-snRNP proteins. For the initial characterization of tri-snRNP proteins that interact directly with U4/U6 snRNPs, immunoaffinity chromatography with an antibody directed against the 60kD protein was performed. We demonstrate that the 60 and 90kD tri-snRNP proteins specifically associate with the U4/U6 snRNP at salt concentrations where the tri-snRNP complex has dissociated. The primary structures of the 60kD and 90kD proteins were determined by cloning and sequencing their respective cDNAs. The U4/U6-60kD protein possesses a C-terminal WD domain that contains seven WD repeats and thus belongs to the WD-protein family, whose best-characterized members include the Gbeta subunits of heterotrimeric G proteins. A database homology search revealed a significant degree of overall homology (57.8% similarity, 33.9% identity) between the human 60kD protein and the Saccharomyces cerevisiae U4/U6 snRNP protein Prp4p. Two additional, previously undetected WD repeats (with seven in total) were also identified in Prp4p, consistent with the possibility that 60kD/Prp4p, like beta-transducin, may adopt a propeller-like structure. The U4/U6-90kD protein was shown to exhibit significant homology, particularly in its C-terminal half, with the S. cerevisiae splicing factor Prp3p, which also associates with the yeast U4/U6 snRNP. Interestingly, U4/U6-90kD shares short regions of homology with E. coli RNase III, including a region encompassing its double-stranded RNA binding domain. Based on their structural similarity with essential splicing factors in yeast, the human U4/U6-60kD and 90kD proteins are likely also to play important roles in the mammalian splicing process.     

烟曲霉几丁质酶基因的克隆与表达

Chi4 4是烟曲霉 (Aspergillusfumigatus)YJ-407产生的一种胞外几丁质酶。通过用真菌几丁质酶保守氨基酸序列与Chi44的N-端序列检索烟曲霉部分基因组序列数据库 ,获得一个编号为contig555的烟曲霉基因组序列 ,可能包含烟曲霉几丁质酶的基因。根据检索结果用RT-PCR方法从烟曲霉YJ-407中克隆到1.4kb的cDNA片段 ,该cDNA的ORF编码一个395个氨基酸的蛋白 ,分子量为43.6kD。对其推导氨基酸序列分析表明该蛋白与其它真菌来源的几丁质酶同源 ,而且活性中心与人巨噬细胞几丁质酶高度同源。该cDNA已在E .coli与PichiapastorisGS115中获得表达 ,分别获得 43kD和44kD的重组蛋白 ,两种重组蛋白均有几丁质酶活性。与野生酶相比 ,大肠杆菌表达的43kD重组酶及Pichia酵母表达的44kD重组酶稳定性下降 ,说明Chi44的糖基化修饰可稳定酶蛋白.     

Escherichia coli cells resistant to the DNA gyrase inhibitor, ciprofloxacin, overproduce a 60 kD protein homologous to GroEL

Using a variety of mutagenic methods, we have generated a series of ciprofloxacin-resistant mutants derived from Escherichia coli strains which overproduce the DNA gyrase A protein. Many of these mutants are found to overexpress a 60 kD protein which is shown to be highly homologous in terms of N-terminal amino acid sequence to the E. coli heat-shock protein, GroEL. Other evidence confirms that the 60 kD protein is unrelated to DNA gyrase and is similar, but not identical, to GroEL.