Identification of the human papilloma virus-1a E4 gene products.

Antibodies prepared against a human papilloma virus-1 (HPV-1) E4/beta-galactosidase fusion protein identified several polypeptides in HPV-1, but not HPV-2 or 4, induced papillomas. The major E4 protein, that represented up to 30% of total cellular protein, was a 16/17-K doublet which was purified by column chromatography and analysed for amino acid content. A peptide derived by chymotryptic digestion was purified by h.p.l.c. and subjected to amino acid sequencing. The unique sequence obtained, Gly-His-Pro-Asp-Leu-Ser-Leu, identified the 16/17-K doublet as a product of the HPV-1 E4 gene region. Antibodies to both the E4/beta-galactosidase fusion protein and the 16/17-K doublet identified two smaller polypeptides (10/11-K) which may represent spliced products of E4. We propose that the products of the HPV-1 E4 gene region are not classical DNA tumor virus early proteins and suggest that they play a role in virus maturation.     

Human Adenovirus Type 2 but Not Adenovirus Type 12 Is Mutagenic at the Hypoxanthine Phosphoribosyltransferase Locus of Cloned Rat Liver Epithelial Cells

Using resistance to the base analog 8-azaguanine as a genetic marker, we showed that adenovirus type 2, but not adenovirus type 12, is mutagenic at the hypoxanthine phosphoribosyltransferase locus of cloned diploid rat liver epithelial cells. Adenovirus type 2 increased the frequency of 8-azaguanine-resistant colonies by up to ninefold over the spontaneous frequency, depending on expression time and virus dose.     

Purification of a serum factor which reverses dibutyryl cAMP induced differentiation

We have previously shown [Grabham et al. (1988) Expt. Eye Res. 47, 123-133] that the adenovirus 12 transformed human retinoblast cell line (Ad 12 HER 10), like a number of other cell types of neuroepithelial origin, can be induced to differentiate in response to exposure to dibutyryl cAMP, and that this differentiation can be rapidly reversed by foetal calf serum. We present data here to show that a single protein, which we have termed differentiation reversal factor (DRF) and have isolated from serum, is responsible for this activity. Following reversal by DRF the growth rate of these cells was shown to be stimulated in serum-free medium. Using ammonium sulphate fractionation, gel filtration chromatography (Ultrogel AcA44), anion exchange chromatography (DEAE cellulose) and preparative gel electrophoresis, DRF has been purified to homogeneity, as judged by polyacrylamide gel electrophoresis in the presence and absence of SDS. The protein has a mol. wt of 72,000 and appears to exist in vivo as a monomer. The concentration of DRF in serum is in the range 100-500 micrograms/ml and is capable of reversing cAMP-induced differentiation of various primary human neuroepithelial cells at physiological concentrations.     

Mutational analysis of human papillomavirus E4 proteins: identification of structural features important in the formation of cytoplasmic E4/cytokeratin networks in epithelial cells.

We have previously demonstrated that human papillomavirus type 1 (HPV 1) and 16 (HPV 16) E4 proteins form cytoplasmic filamentous networks which specifically colocalize with cytokeratin intermediate-filament (IF) networks when expressed in simian virus 40-transformed keratinocytes. The HPV 16 (but not the HPV 1) E4 protein induced the collapse of the cytokeratin networks. (S. Roberts, I. Ashmole, G. D. Johnson, J. W. Kreider, and P. H. Gallimore, Virology 197:176-187, 1993). The mode of interaction of E4 with the cytokeratin IFs is unknown. To identify E4 sequences important in mediating this interaction, we have constructed a large panel of mutant HPV (primarily HPV 1) E4 proteins and expressed them by using the same simian virus 40-epithelial expression system. Mutation of HPV 1 E4 residues 10 to 14 (LLGLL) abrogated the formation of cytoplasmic filamentous networks. This sequence corresponds to a conserved motif, LLXLL, found at the N terminus of other E4 proteins, and similar results were obtained on deletion of the HPV 16 motif, LLKLL (residues 12 to 16). Our findings indicate that this conserved motif is likely to play a central role in the association between E4 and the cytokeratins. An HPV 1 E4 mutant protein containing a deletion of residues 110 to 115 induced the collapse of the cytokeratin IFs in a manner analogous to the HPV 16 E4 protein. The sequence deleted, DLDDFC, is highly conserved between cutaneous E4 proteins. HPV 1 E4 residues 42 to 80, which are rich in charged amino acids, appeared to be important in the cytoplasmic localization of E4. In addition, we have mapped the N-terminal residues of HPV 1 E4 16-kDa and 10/11-kDa polypeptides expressed by using the baculovirus system and shown that they begin at tyrosine 16 and alanine 59, respectively. Similar-sized E4 proteins are also found in vivo. N-terminal deletion proteins, which closely resemble the 16-kDa and 10/11-kDa species, expressed in keratinocytes were both cytoplasmic and nuclear but did not form cytoplasmic filamentous networks. These findings support the postulate that N-terminal proteolytic processing of the E1-- E4 protein may modulate its function in vivo.     

放牧对黄河低阶地盐化草场土壤水盐空间异质性的影响

选定黄河低阶地地形大致相近的放牧草场与封育草场,按实地情况分别设置长770m（放牧）、370m（放牧）和240m（封育）3条样线,采用校正后的便携式WET Sensor以10m为间隔进行土壤水盐的分层（0～10cm,10～20cm）测定,并对其进行地统计学分析.结果表明：黄河低阶地盐化草场表层土壤水盐的变异系数和相关性高于下层,土壤水分的变异系数低于土壤盐分且层间的相关性较高;放牧使土壤水盐之间的相关程度增大,变异降低,且使土壤水分存在一定的冗余.所研究草场的土壤水盐表现为中等或强烈空间自相关,封育盐化草场水盐空间相关尺度的差异较小,而放牧草场则相反.持续放牧已经成为一种稳定影响盐化草场水盐空间分布的结构性因素,使盐化草场的水盐空间异质性减弱.盐生植物在草场内随机分布且不受放牧干扰,是封育盐化草场随机作用较强的最可能原因;而放牧草场内牲畜的啃食和践踏对盐生植物和土壤结构改变所形成的反馈作用大大降低了这些植物对水分特别是盐分的再分配,进而导致放牧草场水盐的含量趋于增大且分布格局趋于简单.     

小麦叶片β-1,3-葡聚糖酶的诱导、纯化与抗菌活性

三个小麦品种331、抗倒680和鲁麦23经氯化汞、水杨酸或核黄素处理后,叶片中的β-1,3-葡聚糖酶活性均有不同程度的升高.氯化汞处理24h对品种331该酶活性的诱导作用最强.因此取用氯化汞处理24 h的331小麦叶片研磨得到粗酶液.将粗酶液经硫酸铵分级沉淀、Phenyl-Sepharose Fast Flow疏水层析、DEAE-Sepharose Fast Flow阴离子交换层析和Sephacryl S-100分子筛层析,得到了SDS-PAGE凝胶电泳谱带单一的β-1,3-葡聚糖酶样品.经SDS-PAGE(12%)和凝胶过滤层析,测得其分子量约为52.0～53.6 kD.抗菌试验测定显示,纯化的β-1,3-葡聚糖酶对供试的4种病原真菌的生长、孢子萌发或芽管伸长都有一定程度的抑制作用.     

人类新基因WDR70的克隆及初步研究

运用基于基因组数据库的特定基因同源新基因的克隆策略得到一个人类新基因WDR70 ,该基因编码一个包含 12个WD4 0结构域的蛋白。WDR70的cDNA序列长 2 2 6 6bp ,预测编码蛋白含 6 30个氨基酸 ,理论分子量为 70× 10 3 u ,染色体定位为 17p13.1。以小鼠胚胎为模型进行整体原位杂交 ,结果显示WDR70基因在 8.5d小鼠胚胎中没有表达 ,而在 9.5d和 10 .5d的小鼠胚胎的脑部有特异表达。由此推断该基因对胚胎期脑部的发育有重要的影响。     

花生幼苗对重复干旱胁迫的生理响应

防雨棚内设盆栽试验,设置对照(Control,75%田间持水量)、干旱胁迫(D,35%)、重复干旱胁迫(D_D,35%)3个处理,探讨花生幼苗对预干旱胁迫的适应和记忆响应,分析预干旱对缓解重复干旱胁迫危害的生理作用。结果表明,与干旱胁迫处理相比,重复干旱胁迫提高了叶片的相对含水量,减少脯氨酸的积累,降低MDA和O·_2~-含量;抗氧化酶SOD、CAT活性降低,其中POD活性降低最为明显,并在复水后恢复到与对照相同水平或低于对照。与正常水分的对照相比,干旱胁迫显著降低叶片光合速率(P_N)、最大光合势能(P_C)、最大光量子产量(Y_Q),但重复干旱处理在重复干旱胁迫时期和复水后P_N、P_C和Y_Q均高于干旱处理。预干旱胁迫导致光合和气孔导度滞后面积、滞后率(H_P和H_g)增加,经过预干旱胁迫后,重复干旱显著降低光合和气孔导度滞后面积和滞后率。预干旱胁迫提高植株在重复干旱胁迫下叶片含水量,减轻重复干旱对植株造成的生理伤害,在光合作用上提高对重复干旱的抵御能力,并在复水后快速恢复到正常水分条件下植株生长水平,减少干旱对植株的不利影响。因此,预干旱胁迫促使花生幼苗具备适应或可记忆初始胁迫的能力,重复干旱胁迫时表现更为迅速和强烈的生理防御和快速的生理恢复机制。     

盐渍化胁迫下油葵对土壤原油污染的适应性及其改良措施

为了探究植物在盐渍化胁迫下对原油污染的适应性及改良措施,本研究以油葵作为研究对象,进行了原油-氯化钠-脱硫石膏盆栽正交试验和煤渣-沸石-脱硫石膏-锯沫盆栽正交试验.结果表明: 在盐渍化条件下,随着原油浓度的增大,油葵幼苗株高相对生长率(RGR)、地上生物量RGR、根氮磷比均显著减小,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性呈先增加后显著降低的趋势;随着锯沫体积分数的加大,油葵株高RGR和地上生物量RGR均显著增加,SOD活性逐渐降低,说明锯沫在改良盐渍化原油污染土壤方面比煤渣、沸石和脱硫石膏效果显著.在盐渍化条件下,原油污染能够降低油葵幼苗的生长率,锯沫对改良原油污染有较好的效果.     

甘肃省民勤沙区土壤结皮理化性质研究

以甘肃民勤沙区为研究区域 ,分别采集了不同地貌部位和不同植被类型下的土壤结皮 ,并对其理化性质进行了初步分析。从结皮土壤的机械组成、土壤盐分、土壤养分、土壤阳离子交换量等方面来看 ,丘间地状况都明显要优于灌丛沙包。这可能与丘间地地形低洼 ,有利于土壤物质汇聚 ,以及其接受的大气降尘远较灌丛沙包为多有关。对于灌丛沙包来讲 ,白刺沙包在理化性质上 ,其状况要优于红柳结皮和梭梭结皮 ,这主要与其对环境的适应性及其所处的演替阶段有关。从目前的植被演替情况来看 ,白刺是当地的顶极种群 ,最适应环境 ,因此结皮发育状况也好 ;红柳目前已经处于极度退化进程中 ,而梭梭为人工植被 ,人工植被因在演替阶段中不起决定作用 ,故理化性质较差。另外从该研究工作还可以看出 ,对白刺植被采取围封措施之后 ,可以显著地促进结皮的生长发育 ,提高结皮中的有机质、全 N、全 P、速效 N等养分以及 Ca CO3含量。