大肠癌中p53基因突变的研究

应用聚合酶链反应(PCR)──单链构型多态性(SSCP)结合银染法对14例大肠癌p53基因的第4、第5─6和第7外显子进行了点突变的研究,结果共检测出6例点突变,而且发现各外显子的突变频率存在差异。另外,利用购自ATCC的两个探针 (p53cDNA探针和pYNZ22探针)对大肠癌中p53基因的杂合性失去进行了研究,在14例大肠癌中共检出6例杂合性丢失。将点突变检测结果同杂合性丢失结果进行比较分析, 并着重探讨了大肠癌中p53基因失活导致肿瘤的作用方式。 Abstract:The exons 4-7 of p53 gene were examined in 14 colorectal Cancer patients by using PCR-SSCP-silver staining method.The results showed 6 cases of point mutation and the mutation frequencies of exons were different from each other.p53 cDNA and pYNZ22 VNTR were used as probes to examine LOH(Loss of heterozygosity)of 14 colorectal cancers.6 cases with LOH were found.The results of present research suggest that mutation and LOH of p53 gene are critical events in the progress and development of Cancer.There were different kinds of inactivation model of p53 gene in the process of development of cancer and transformation of cells.     

腺苷三磷酸和环腺苷单磷酸对丝状真菌纤维素酶合成的调节

以虫荧光素酶法检验了四株丝状真菌在葡萄糖—无机盐液体培养过程中的胞内ATP含量。结果表明,只有当胞内ATP浓度低于10~(-S)mg/ml时,真菌才开始合成胞外纤维素酶(FPA)。以不同浓度的各种碳源培养时,菌体胞内ATP含量只要超过10~(-1)mg/ml,FPA的合成即发生阻遏。菌体胞内ATP含量与FPA合成呈显著负相关。以高效液相色谱(HPLC)法检测了菌体培养液中的cAMP含量。在非阻遏条件下,外源cAMP可以提高FPA的合成水平。但外源cAMP不能解除已经发生的酶合成阻遏。菌体ATP和cAMP水平是调节真菌纤维素酶合成的重要因子。     

L-山梨糖提高木霉纤维素酶合成速率机制的研究

在0.5％葡萄糖-Mandels盐培养液中添加终浓度为0.5％的L-山梨糖,可使里斯木霉(Trichoderma reesei C 30)和拟康氏木霉(Trichoderma pseudokoningii S38)的内切和外切β-1,4葡聚糖酶合成速率在培养的2天内提高4倍。与此同时β-葡萄糖苷酶的合成没有明显变化。L-山梨糖能明显抑制菌丝生长,但对葡萄糖的吸收没有影响,对菌丝分泌纤维素酶的机制影响不大。其对酶合成的促进可能主要是通过降低了菌丝体的生长速率。     

人粒细胞集落刺激因子（hG－CSF）cDNA3′－UTR对其表达的影响

利用人粒细胞集落刺激因子（ｈＧ－ＣＳＦ）ｃＤＮＡ３′端非翻译区（３′－ＵＴＲ）中存在的ＤｒａⅠ酶切位点,通过部分酶切与完全酶切,删除３′－ＵＴＲ不同长度,构建了四种ｈＧ－ＣＳＦｃＤＮＡ瞬时重组表达质粒。转染ＣＯＳ－７细胞后,生物活性测定结果提示,ｈＧ－ＣＳＦｃＤＮＡ３′－ＵＴＲ对其表达起负调控作用,其关键性序列位于紧接终止密码子ＴＧＡ下游的６５ｂｐ范围内,３′－ＵＴＲ对ｈＧ－ＣＳＦｃＤＮＡ表达的影响与转录水平的差别有一定关系。     

细胞色素c在氨基酸及二肽修饰金电极上的电化学反应

系统地研究了细胞色素ｃ在多种氨基酸和多肽修饰电极上的电化学反应。并对影响加速细胞色素ｃ电化学反应的因素进行了讨论。     

Leaf and canopy photosynthetic CO2 uptake of a stand of Echinochloa polystachya on the Central Amazon floodplain

The C₄ grass Echinochloa polystachya, which forms dense and extensive monotypic stands on the Varzea floodplains of the Amazon region, provides the most productive natural higher plant communities known. The seasonal cycle of growth of this plant is closely linked to the annual rise and fall of water level over the floodplain surface. Diurnal cycles of leaf photosynthesis and transpiration were measured at monthly intervals, in parallel with measurements of leaf area index, canopy light interception and biomass. By artificial manipulation of the light flux incident on leaves in the field light-response curves of photosynthesis at the top and near to the base of the canopy were generated. Fitted light-response curves of CO₂ uptake were combined with information of leaf area index, incident light and light penetration of the canopy to estimate canopy rates of photosynthesis. Throughout the period in which the floodplains were submerged photosynthetic rates of CO₂ uptake (A) for the emergent leaves were high with a mean of c. 30 mol m^-2 s^-1 at mid-day and occasional values of 40 mol m^-2 s^-1. During the brief dry phase, when the floodplain surface is uncovered, there was a significant depression of A, with mid-day mean values of c. 17 mol m^-2 s^-1. This corresponded with a c. 50% decrease in stomatal conductance, and a c. 35% depression in the ratio of the leaf inter-cellular to external CO₂ concentration (c _i/c _a). During the dry phase, a midday depression of rates of CO₂ assimilation was observed. The lowest leaf area index (F) was c. 2 in November–December, when the flood plain was dry, and again in May, when the rising floodwaters were submerging leaves faster than they were replaced. The maximum F of c. 5 was in August when the floodwaters were receding rapidly. Canopy light interception efficiency varied from 0.90 to 0.98. Calculated rates of canopy photosynthesis exceeded 18 mol C m^-2 mo^-1 throughout the period of flooding, with a peak of 37 mol C m^-2 mo^-1 in August, but declined to 13 mol C m^-2 mo^-1 in November during the dry phase. Estimated uptake of carbon by the canopy from the atmosphere, over 12 months, was 3.57 kg C m^-2. This was insufficient to account for the 3.99 kg C m^-2 of net primary production, measured simultaneously by destructive harvesting. It is postulated that this discrepancy might be accounted for by internal diffusion of CO₂ from the CO₂-rich waters and sediments via the roots and stems to the sites of assimilation in the leaves.     

花粉高尔基囊泡与牛脑微管的体外结合

把从榛木（Ｃｏｒｙｌｕｓａｖｅｌｌａｎａ Ｌ．）花粉中分离得到的高尔基囊泡与经高度纯化并聚合好的牛脑微管进行体外组合,然后于１．５ ｍ ｏｌ／Ｌ的蔗糖层上进行超离心,对其沉淀物进行ＳＤＳ－聚丙烯酰胺凝胶电泳和电镜负染。结果表明,花粉高尔基囊泡可以结合到牛脑微管上,证明植物花粉的高尔基囊泡与动物细胞的某些细胞器一样,也与细胞骨架的主要组成之一——微管具有结构上的紧密联系。花粉高尔基囊泡与牛脑微管的体外结合能力,受１０ ｍ ｍ ｏｌ／ＬＡＴＰ和０．５ ｍ ｏｌ／ＬＫＣｌ的影响,但不受５ ｍ ｍ ｏｌ／Ｌ ＡＭＰ－ＰＮＰ的影响,说明两者结合可能是通过高尔基囊泡表面与ＡＴＰ有关的某种外周膜蛋白来完成的。     

Analysis of the frequency of inheritance of transposed Ds elements in Arabidopsis after activation by a CaMV 35S promoter fusion to the Ac transposase gene

The Ac/Ds transposon system of maize shows low activity in Arabidopsis. However, fusion of the CaMV 35S promoter to the transposase gene (35S::TPase) increases the abundance of the single Ac mRNA encoded by Ac and increases the frequency of Ds excision. In the experiments reported here it is examined whether this high excision frequency is associated with efficient re-insertion of the transposon. This was measured by using a Ds that carried a hygromycin resistance gene (HPT) and was inserted within a streptomycin resistance gene (SPT). Excision of Ds therefore gives rise to streptomycin resistance, while hygromycin resistance is associated with the presence of a transposed Ds or with retention of the element at its original location. Self-fertilisation of most individuals heterozygous for Ds and 35S::TPase produced many streptomycin-resistant (strep^r) progeny, but in many of these families a small proportion of strep^r seedlings were also resistant to hygromycin (hyg^r). Nevertheless, 70% of families tested did give rise to at least one strep^r, hyg^r seedling, and over 90% of these individuals carried a transposed Ds. In contrast, the Ac promoter fusion to the transposase gene (Ac::TPase) produced fewer strep^rhyg^r progeny, and only 53% of these carried a transposed Ds. However, a higher proportion of the strep^r seedlings were also hyg^r than after activation by 35S::TPase. We also examined the genotype of strep^r, hyg^r seedlings and demonstrated that after activation by 35S::TPase many of these were homozygous for the transposed Ds, while this did not occur after activation by Ac::TPase. From these and other data we conclude that excisions driven by 35S::TPase usually occur prior to floral development, and that although a low proportion of strep^r progeny plants inherit a transposed Ds, those that do can be efficiently selected with an antibiotic resistance gene contained within the element. Our data have important implications for transposon tagging strategies in transgenic plants and these are discussed.