转铁蛋白结构与功能的研究——骆驼血清转铁蛋自的分离纯化及其含单一铁结合部位的结构域的制备和鉴定

分离纯化获得的骆驼血清转铁蛋白由分子量为73,000和63,000两个组分组成。两者至少N-端五肽顺序相同(Met-Pro-Asp-Lys-Thr)。骆驼血清转铁蛋白在生理pH下不能与人胎盘转铁蛋白受体结合。用胰蛋白酶酶解骆驼转铁蛋白可以同时得到两个合单一铁结合部位的结构域,分别来自转铁蛋白分子的N-端称N-端结构域(分子量34,700和40,700)和C-端称C-端结构域(分子量35,100)。在上述结果的基础上指出并讨论了反刍动物转铁蛋白在结构和功能上存在更多的共同性,而与其它哺乳动物的转铁蛋白有着明显的区别。     

Making transgenic livestock: genetic engineering on a large scale.

The feasibility of introducing foreign genes into the genomes of cattle, goats, pigs, and sheep has only recently been demonstrated. Studies have thus far focused on improving growth efficiency or directing expression of pharmaceutical proteins to the mammary glands of these species. The general strategy for producing transgenic livestock and mice is similar. In addition to the obvious difference in scale between mice and livestock experiments, there are noteworthy obstacles that significantly reduce the efficiency of producing transgenic livestock. Low embryo viability, low transgene integration rates, and high animal costs contribute to project costs that can easily exceed hundreds of thousands of dollars. A better understanding of the mechanisms that govern transgene integration should lead to improved efficiencies. But, the full potential of the transgenic livestock system will not be fully realized until: 1) gene constructs can be designed that function in a reproducible, predictable manner; and 2) the genetic control of physiological processes are more clearly elucidated. Newly emerging approaches may resolve at least some of these issues within the next decade.     

Pea lectin is correctly processed,stable and active in leaves of transgenic potato plants

A gene encoding the preproprotein of the pea (Pisum sativum) lectin was expressed in transgenic potato plants using a cauliflower mosaic virus (CaMV) 35S promoter or a tobacco ribulose bisphosphate carboxylase small subunit (ssRubisco) promoter. Presence of the pea lectin to levels greater than 1% of total soluble leaf protein was detected by radioimmunoassay (RIA). The pattern of expression derived from the two promoters was established using both RIA and a squash-blot immunolocalisation technique. Western blotting demonstrated that the preproprotein was correctly processed, generating and subunits that assembled to give an isolectin form observed in pea seeds and roots. It was also found that the haemagglutination activity and specificity of pea lectin synthesised in transgenic potato leaves was comparable to purified lectin from pea cotyledons.     

Phenotypically normal transgenic T-cyt tobacco plants as a model for the investigation of plant gene expression in response to phytohormonal stress

The tumour-inducing T-DNA gene 4 (T-cyt gene) of the nopaline Ti plasmid pTiC58 was cloned and introduced into tobacco cells by leaf disc transformation using Agrobacterium plasmid vectors. Tobacco shoots exposed to elevated cytokinin levels were unable to develop roots and lacked apical dominance. Using exogenously applied phytohormone manipulations we were able to regenerate morphologically normal transgenic tobacco plants which differed in endogenous cytokinin levels from normal untransformed plants. Although T-cyt gene mRNA levels, as revealed by dot-blot hybridization data, in these rooting plants were only about half those in primary transformed shoots the total amount of cytokinins was much lower than in crown gall tissue or cytokinin-type transformed shoots as reported by others. Nevertheless the cytokinin content in T-cyt plants was about 3 times greater than in control tobacco plants.Elevated cytokinin levels have been shown to change the expression of several plant genes, including some nuclear genes encoding chloroplast proteins. Our results show that the mRNA levels of chloroplast rbcL gene increase in cytokinin-type transgenic tobacco plants as compared with untransformed plants. Data obtained suggest that T-cyt transgenic plants are a good model for studying plant gene activity in different parts of the plant under endogenous cytokinin stress.     

Differential inactivation and methylation of a transgene in plants by two suppressor loci containing homologous sequences

In a previous study on doubly transformed tobacco plants, we observed the unexpected inactivation in trans of T-DNA-I (encoding Kan^rNOS) following the introduction into the same genome of an unlinked copy of T-DNA-II (encoding Hyg^rOCS). This inactivation, which probably resulted from interactions between homologous regions on each T-DNA, was correlated with methylation in the nos _pro, which controlled the expression of both the nptII and nos genes. In this paper, we show that the inactivation and methylation of the nos _pro nptII gene in the presence of a suppressor T-DNA-II locus can be either complete (epistasis) or partial (cellular mosaicism). In plants showing partial suppression, the strength of the Kan^r phenotype, which apparently reflected the proportion of cells expressing the nptII gene, was inversely correlated with the degree of methylation of the nos _pro. The extent of nos _pro methylation decreased progressively in successive generations as suppressor T-DNA-II loci were crossed out. The strength of the Kan^r phenotype was improved and nos _pro methylation was less extensive in first generation Kan^r progeny obtained from outcrossing with untransformed tobacco than from self-fertilization.     

Mechanism of autoxidation of oxyhaemoglobin

Oxyhaemoglobins from erythrocytes of different animals including fish, amphibians, reptiles, birds, mammals and human beings have been isolated by ion-exchange chromatography over phosphocellulose and the comparative rates of autoxidation of oxyhaemoglobin studied. The mechanism of autoxidationin vitro has been elucidated using toad as well as human oxyhaemoglobin. Autoxidation is markedly inhibited by carbon monoxide as well as by anion ligands, namely, potassium cyanide, sodium azide and potassium thiocyanate. The inhibition by anions is in the same order as their strength as nucleophiles, indicating that it is the oxyhaemoglobin and not the ligand-bound deoxy species which undergoes autoxidation. The structure of oxyhaemoglobin is considered to be mainly and determination of the rate of autoxidation with or without using superoxide dismutase and catalase indicates that the initial process of autoxidation takes place by dissociation of to methaemoglobin and superoxide to the extent of 24%. The superoxide thus produced reattacks oxyhaemoglobin to produce further methaemoglobin and hydrogen peroxide. H₂O₂ is a major oxidant of oxyhaemoglobin producing methaemoglobin to the extent of 53%. A tentative mechanism of autoxidation showing the sequence of reactions involving superoxide, H₂O₂ and OH has been presented.     

大熊猫(Ailuropoda melanolenca)显带染色体的研究

大熊猫系我国特产的世界珍稀动物,素有“活化石”和“国宝”之称。限于材料来源,虽有核型的少数报道（邓承宗等,1980;陈文元等,1984;Newnham et al.,1966）,但研究尚不够深入。1980年,Wurster-Hill和Bush首先报道了大熊猫（）的显带核型,并与杂交熊等比较,探讨了大熊猫的分类地位。本文对四只大熊猫的G带、C带核型和Ag-NORs作了分析,绘制了G带核型模式图,并提出了某些商榷的意见。     

Standardization of Radiorenography in Dehydrated and Rehydrated Primates Under Laboratory Conditions

Radiorenography with ^99mTo-labelled diethylenctriaminepcntacetic acid ([^99mTc]-DTPA) was performed on chacma baboons (Papio ursinus) and vervet monkeys (Cereopithecus pygerythus) to establish the effects of various states of hydration on the data obtained from the DTPA-renogram. The renogram parameters, which can be related to certain aspects of kidney function, varied significantly with the degree of hydration. It is therefore imperative for clinically directed animal research projects on the urinary system to standardise the experimental procedure for radiorenography. A dehydration of 6 h followed by an hour IU rehydration period using 200 ml of a 0.9% NaCI solution on baboons under thiopentone sodium anaesthetic, was found to be the most suitable procedure for radiorenographic investigations in this primate model.     

In Vivo Release from Cerebral Cortex of [14C]Glutamate Synthesized from [U-14C]Glutamine

Awake, unrestrained, and behaviourally normal animals with superfusion cannulae implanted over the sensorimotor cortex were used in a study of the capacity of infused [U-14C]glutamine for labelling glutamate and other amino acids released by depolarising stimuli. A spontaneous background release of [14C]glutamate was detected. This was increased by tityustoxin (1 microM). The specific radioactivity of glutamate increased eightfold during the evoked-release period. [14C]Aspartate was also detected and showed increased release, but not increased specific labelling, in response to depolarisation. Evoked gamma-aminobutyric acid (GABA) release occurred but only small amounts of [14C]GABA were detected. Glutamine showed increased rates of uptake to the sensorimotor cortex during stimulation periods, suggesting an accelerated breakdown via glutaminase.