抗冻蛋白研究进展

南北两极的鱼类生活在低于 0℃ (约- 1 .9℃ )的海水中 ,适应于这种环境 ,其体液内有抗冻物质———抗冻蛋白 (ＡＦＰ)或抗冻糖蛋白 (ＡＦＧＰ) ,以防止体液内冰核的形成与生长。在越冬昆虫体内 ,有活性更高的抗冻蛋白。近年来在耐寒植物中也陆续发现了抗冻蛋白。本文将介绍各类抗冻蛋白的结构和生化性质、功能特性、抗冻作用机制、有关抗冻蛋白基因工程研究及抗冻蛋白的应用研究。1 .抗冻蛋白分子结构及生化特性1 .1　ＡＦＧＰ　　ＡＦＧＰ肽链是由Ａｌａ Ａｌａ Ｔｈｒ三肽单位重复组成 ,苏氨酸残基上接双糖基团 [3 Ｏ ( β Ｄ 半乳糖 ) …     

植物细胞质膜H+-ATPase的结构与功能

植物细胞质膜Ｈ＋ＡＴＰａｓｅ属于Ｐ型质子泵。由该酶产生的跨膜电化学梯度是物质跨膜运输的原初动力。研究表明,质膜Ｈ＋ＡＴＰａｓｅ与植物的生长发育密切相关,被称为植物细胞的“主宰酶”。近年,关于该酶的生化特性,基因表达与调控以及结构与功能等方面的研究取得重要进展。对质膜Ｈ＋ＡＴＰａｓｅ的生化特性,分子结构,调节机制和生理功能等进行了综述     

真核生物tRNA基因的表达

真核生物ｔＲＮＡ基因的表达金由辛（中科院上海生化所分子生物学国家重点实验室,上海２０００３１）真核细胞的生长周期真核生物ｔＲＮＡ基因的表达研究远落后于蛋白质基因的表达研究。原因很明显：１。ｔＲ－ＮＡ基因表达的终产物为ＲＮＡ,它在中心法则遗传信息传递过程中才走了半程;2。蛋白质基因表达受基因工程的推动。     

大熊猫生理生化及细胞遗传学研究与进展

大熊猫生理生化及细胞遗传学研究与进展彭建军石红艳（四川师范学院珍稀动植物研究所南充６３７００２）关键词大熊猫生理生化细胞遗传自１８６９年大熊猫（Ａｉｌｕｒｏｐｌｏｄａｍｅｌａｎｏｌｅｕｃａ）得以订名以来,由于它的古老、珍稀、奇特和狭窄分布,一直吸引...     

试管苗玻璃化现象的生理生化和机理探讨

试管苗玻璃化现象的生理生化和机理探讨梁海曼,周菊华（杭州大学生物系杭州３１００１２）关键词试管苗,玻璃化现象,生理生化特点,机理ＰＨＹＳＩＯＬＯＧＩＣＡＬ,ＢＩＯＣＨＥＭＩＣＡＬＣＨＡＲＡＣＴＥＲＩＳＴＩＣＳＡＮＤＭＥＣＨＡＮＩＳＭＥＸＰＬＯＲＡＴＩ...     

基因表达的转录后与翻译起始调控

基因表达的转录后与翻译起始调控彭晓冬,童坦君（北京医科大学生化与分子生物学系,北京１０００８３）关键词转录后调控,翻译起始调控,ＲＮＡ结构近年来,基因表达的转录调控研究十分深入。转录以后诸过程的调控机制研究亦越来越引人注目。一般认为,对ｍＲＮＡ成熟、...     

低浓度SO2对云杉幼苗中碳水化合物代谢的影响研究

研究了低浓度ＳＯ２对云杉幼苗的光合作用速率及碳水化合物代谢的生理生化过程的影响,测定了ＣＯ２摄取速率、幼苗蔗糖含量、蔗糖磷酸合成酶、蔗糖合成酶和转化酶活性及ＡＴＰ／ＡＤＰ值．结果表明,低浓度的ＳＯ２（２００μｇ·ｍ（－３））能瞬时刺激幼苗对ＣＯ２的吸收．与对照组相比,幼苗中的蔗糖含量及蔗糖磷酸酶活性明显下降,ＡＴＰ／ＡＤＰ处于低水平．因此,在受到ＳＯ２污染形成可见伤害前,幼苗的生理生化过程可能已受到干扰．     

桃儿七生化生态适应的初步研究

桃儿七生化生态适应的初步研究马绍宾胡志浩李俊（云南大学生物系,昆明６５００９１）（云南省香料中心,昆明６５００５１）ＡＰｒｉｌｉｍｉｎａｒｙＳｔｕｄｙｏｆｔｈｅＢｉｏｃｈｅｍｉｃａｌＥｃｏｌｏｇｉｃａｌＡｄａｐｔａｔｉｏｎｏｆＳｉｎｏｐｏｄｏｐｈｙ...     

本刊顾问刘新垣院士简介

本刊顾问刘新垣院士简介中科院院士,上海生化所研究员,上海华新生物高技术有限公司董事长刘新垣教授在分子生物学领域中取得许多重要的研究成果。他６０年代进行了ＲＮＡ结构和功能的研究,改进了ＲＮＡ酶解指纹图谱分析方法,并首次证实丝腺大分子ＲＮＡ中含稀有碱基,...     

V型H~＋_－ATP酶的分子结构及其药理学意义

Ｖ型ＡＴＰ酶广泛存在于细胞内膜系统,如溶酶体,内膜体,高尔基体,分泌颗粒等,Ｖ－ＡＴＰ酶水解ＡＴＰ,建立跨膜质子电化学梯度（△￣μＨ￣＋）,酸化细胞内外环境。研究证明△￣μＨ￣＋和酸化作用为细胞的内吞、外泌、膜流和物质转运等生理生化反应提供了必需的条件。Ｖ－ＡＴＰ酶在生命活动中的重要性和它的实际意义,日益引起人们的兴趣与关注,是当前Ｈ￣－ＡＴＰ酶家族中一个异常活跃的研究分支。     

Regulation of gene expression in Salmonella phage P22. II. Regulation of expression of late functions

Transactivation experiments were performed involving the genetically related Salmonella phages P22, L and Px1 in order to find out if more than one positively acting regulatory product is engaged in the expression of vegetative gene functions of each of these phages. The results obtained with Px1- and L-lysogenic cells superinfected with P22 suggest the following conclusions: 1. The expression of the early genes 12 and 23 and of the late gene 19 (lysozyme synthesis) is positively regulated by two different regulatory products, since P22 transactivates in prophage Px1 both early and late genes (Prell, 1973), in prophage L only late genes. 2. The transactivation by P22 of the lysozyme gene of prophage L takes place in the presence of L repressor. This conclusion is suggested, since the superinfecting P22 does not derepress early gene expression (see 1.), and is confirmed by demonstration of replication inhibition for L phage in L lysogenic cells doubly superinfected with L and P22 phages (Thomas-Bertani-experiment). 3. The late gene regulatory protein seems to be synthesized by gene 23, as transactivation experiments with both L- and Px1 prophages suggest. 4. The expression of gene 23 itself is turned on by an early regulatory product. The gene which codes for it is still unidentified. However its product seems to by highly specific, since it is active on Px1- but not on L-prophage.     

An HSV-1 Vector Containing the Rat Tyrosine Hydroxylase Promoter Enhances Both Long-Term and Cell Type-Specific Expression in the Midbrain

Abstract: A defective herpes simplex virus type one (HSV-1) vector that contains a 6.8-kb fragment of the rat tyrosine hydroxylase promoter (pTHlac-7kb) was examined for its capability to target catecholaminergic cell type-specific expression in the CNS. Cell type-specific expression was assessed by comparison with a control vector (pHSVlac) that uses the HSV-1 immediate early 4/5 promoter to support expression in multiple cell types. In initial experiments comparing expression in catecholaminergic and noncatecholaminergic cell lines, pTHlac-7kb supported a seven- to 20-fold increase in reporter gene expression in catecholaminergic cell lines. Four days after stereotactic injection into the midbrain of adult rats, pTHlac-7kb supported a 10-fold targeting of β-galactosidase expression to tyrosine hydroxylase-expressing neurons in the substantia nigra pars compacta compared with pHSVlac. Expression from pTHlac-7kb was stably maintained for 6 weeks with no significant changes in the pattern of expression. Long-term expression from pTHlac-7kb was confirmed by RNA and DNA analysis. In contrast, reporter gene expression in the midbrain from pHSVlac decreased ∼30-fold between 4 days and 6 weeks after gene transfer. Thus, within the context of this HSV-1 vector system, the tyrosine hydroxylase promoter enhanced cell type-specific expression and contributed to stable, long-term expression of a recombinant gene product in neurons. The capability to target recombinant gene expression to catecholaminergic neurons in specific brain areas may be useful for studies on the roles of these neurons in brain physiology and behavior.     

Structure and myofiber-specific expression of the rat muscle regulatory gene MRF4.

We have cloned an 11.3-kb rat genomic DNA fragment encompassing the muscle regulatory factor 4 (MRF4) protein-coding sequence, 8.5 kb of 5'-flanking sequence, and 1.0 kb of 3'-flanking sequence. In order to study MRF4 gene expression, the rat myogenic cell line, L6J1-C, which expresses the endogenous MRF4 gene only in differentiated myofibers, was transfected stably with the full-length genomic clone and various 5' deletions. RNase protection assays demonstrated that MRF4 genes containing as little as 430 bp of 5'-flanking sequence exhibited an increase in expression as the cells differentiated into myofibers, indicating that elements responsible for fiber-specific expression are contained within this cloned DNA fragment. Similar up-regulation was observed with genes containing 1.5 kb of 5'-flanking sequence. Interestingly, MRF4 genes containing 5.0 kb and 8.5 kb of 5'-flanking sequence were up-regulated to even higher levels, suggesting that additional myofiber-specific regulatory elements located between 1.5 and 5.0 kb upstream from the coding region play a role in regulating the expression of this muscle-specific gene.     

A target-specific feeding toxicity of β(1) integrin dsRNA against diamondback moth, Plutella xylostella

Integrin is a cell-surface protein consisting of α and β heterodimers. A predicted amino acid sequence of an integrin subunit of the diamondback moth, Plutella xylostella, was highly homologous to other lepidopteran β1 subunits and possessed essential functional domains. The β1 integrin of P. xylostella (βPx1) was expressed in all developmental stages of P. xylostella. It was also expressed in all tested tissues including hemocyte, fat body, gut, and epidermis of last instar. When βPx1 expression was suppressed by injection of dsRNA specific to βPx1 (dsRNA(βPx1)), the treated larvae exhibited significant suppression in immune response and also suffered significant larval mortality. When dsRNA(βPx1) was orally fed to young larvae, it suppressed the expression of aPx1 and resulted in a significant mortality. By contrast, a dsRNA specific to β1 subunit of Spodoptera exigua gave little adverse effects on βPx1 expression and larval development when it was treated by injection or oral administration, though these two genes showed 71% sequence homology. These results suggest a target-specific RNA interference of dsRNA(βPx1), which causes significant mortality to P. xylostella by feeding treatment.     

Distal and proximal cis-linked sequences are needed for the total expression phenotype of the mouse alcohol dehydrogenase 1 (Adh1) gene

Mouse alcohol dehydrogenase 1 (Adh1) gene expression occurs at high levels in liver and adrenal, moderate levels in kidney and intestine, low levels in a number of other tissues, and is undetectable in thymus, spleen and brain by Northern analysis. In transgenic mice, a minigene construct containing 10 kb of upstream and 1.5 kb of downstream flanking sequence directs expression in kidney, adrenal, lung, epididymis, ovary and skin but promotes ectopic expression in thymus and spleen while failing to control expression in liver, eye, intestine and seminal vesicle. Cosmids containing either 7 kb of upstream and 21 kb of downstream or 12 kb of upstream and 23 kb of downstream sequence flanking genetically marked Adh1 additionally promotes seminal vesicle expression suggesting downstream or intragenic sequence controls expression in this tissue. However, expression in liver, adrenal, or intestine is not promoted. The Adh1(a) allele on the cosmid expresses an enzyme electrophoretically distinct from that of the endogenous Adh1(b) allele, and presence of the heterodimeric enzyme in expressing tissues confirms that transgene activity occurs in the same cell-type as the endogenous gene. Transgene expression levels promoted by cosmids were at physiologically relevant amounts and exhibited greater copy-number dependence than observed with minigenes. Transgene mRNA expression correlated with expression measured at the enzyme level. A bacterial artificial chromosome containing 110 kb of 5'- and 104 kb of 3'-flanking sequence surrounding the Adh1 gene promoted expression in tissues at levels comparable to the endogenous gene most importantly including liver, adrenal and intestinal tissue where high level Adh1 expression occurs. Transgene expression in liver was in the same cell types as promoted by the endogenous gene. Although proximal elements extending 12 kb upstream and 23 kb downstream of the Adh1 gene promote expression at physiologically relevant levels in most tissues, more distal elements are additionally required to promote normal expression levels in liver, adrenal and intestinal tissue where Adh1 is most highly expressed.