RT－cDNA克隆－PCR法获取犬瘟热病毒融合蛋白基因（F）

纯化鸡胚成纤维细胞培养的犬瘟热病毒（ＣａｎｉｎｅＤｉｓｔｅｍｐｅｒＶｉｒｕｓ,ＣＤＶ）,获得病毒基因组ＲＮＡ后,反转录合成双链病毒Ｆ基因ｃＤＮＡ。将此双链ｃＤＮＡ平端插入ＰＵＣ１９质粒ＳａｍⅠ位点构建重组质粒,进行ｃＤＮＡ克隆。以重组克隆质粒为模板ＰＣＲ扩增,获得ＣＤＶ全长Ｆ基因。将此Ｆ基因插入表达载体ＰＢＶ２２０,在大肠杆菌中表达,通过对表达产物的最终鉴定,可确认所获片段为ＣＤＶ全长Ｆ基因．     

GM_3对小鼠腹腔巨噬细胞磷脂代谢转换率的影响

神经节苷脂ＧＭ_３对小鼠腹腔常驻巨噬细胞（Ｒ－Ｍ）和Ｇｅ－１３２体内激活的巨噬细胞（Ｇｅ－１３２－Ｍ）的磷脂代谢转换有显著的影响,当这两种Ｍ在体外用ＧＭ_３处理时,表现出［￣（３２）Ｐ］Ｐｉ和［￣３Ｈ］肌醇参入ＰＩ降低,参入ＰＩＰ、ＰＩＰ_２增加;但在［￣（３２）Ｐ］Ｐｉ和［￣３Ｈ］胆碱参入ＰＣ上,Ｒ－Ｍ与Ｇｅ－１３２－Ｍ不同,即ＧＭ_３促进同位素前体参入Ｒ－Ｍ的ＰＣ,抑制它们参入Ｇｅ－１３２－Ｍ的ＰＣ．以上结果表明ＧＭ３可能提高了ＰＩ或ＰＩＰ的磷酸激酶的活性,致使［￣（３２）Ｐ］ＰＩＰ和［￣（３２）Ｐ］ＰＩＰ_２增多,［￣（３２）Ｐ］ＰＩ减少．激活的Ｍ（Ｇｅ－１３２－Ｍ）本身ＰＣ代谢转换率较Ｒ－Ｍ高,当Ｇｅ－１３２－Ｍ再受ＧＭ_３刺激,ＰＣ代谢转换率降低,这提示ＧＭ_３对激活的Ｍ的ＰＣ代谢转换有调节作用．     

Yeast colonizing the intestine of rainbow trout (Salmo gairdneri) and turbot (Scophtalmus maximus)

Yeast were isolated from the intestine of farmed rainbow trout (Salmo gairdneri), turbot (Scophtalmus maximus), and free-living flat-fish (Pleuronectes platessa and P. flesus). The average number of viable yeasts recovered from farmed rainbow trout was 3.0 × 10³ and 0.5 × 10² cells per gram homogenized intestine for white and red-pigmented yeasts, respectively. The dominant species were Debaryomyces hansenii, Saccharomyces cerevisiae, Rhodotorula rubra, and R. glutinis. In 5 of 10 free-lving marine fish, > 100 viable yeast cells per gram intestinal mucus were recovered. Red-pigmented yeasts dominated and composed >90% of the isolates. Colonization experiments were performed by inoculating rainbow trout and turbot with fish-specific, isolated yeast strains and by examining the microbial intestinal colonization at intervals. Inoculation of experimental fish with pure cultures of R. glutinis and D. hansenii HF1 yielded colonization at a level several orders of magnitude higher than before the inoculation. Up to 3.8 × 10⁴, 3.1 × 10⁶, and 2.3 × 10⁹ viable yeast cells per gram intestine or feces were recovered in three separate colonization experiments. The high level of colonizing yeasts persisted for several weeks. The concentrations of yeasts in the tank water never exceeded 10³ viable cells per milliliter. No traces of fish sickness as a result of high yeast colonization were recorded during any of the colonization experiments. For periods of the experiments, the concentration of aerobic bacteria in the fish intestine was lower than the intestinal yeast concentration. Scanning electron microscopy studies demonstrated a close association of the yeasts with the intestinal mucosa. The mucosal colonization was further demonstrated by separating intestinal content, mucus, and tissue. All compartments were colonized by >10³ viable yeast cells per gram. No bacteria were detected on the micrographs, indicating that their affinity for the intestinal mucosa was less than that of the yeasts. Correspondence to: Thomas Andtid     

RFLP tagging of a salt tolerance gene in rice

A salt tolerant rice mutant (M-20) was obtained through selection in vitro. Its tolerance was stably inherited over eight generations and most traints between M-20 and its sensitive original 77–170 (Oryza sativa) were very similar. By deriving an F₂ population of M-20 × 77–170 and splitting every F₂ individual into two parts, with one part planted in normal conditions and another part in saline conditions, the inheritance of salt tolerance in rice was studied. Under normal conditions, there was no apparent segregation among F₂ individuals. Under saline conditions, however, the segregation of traits was obvious. According to our standards, the ratio of salt sensitive:moderately-tolerant:tolerant plants was 25:42:18, in accordance with a 1:2:1 ratio. It suggested that the improvement of salt tolerance in our materials was induced by the mutation of a major tolerant gene which showed incomplete dominance. By use of 130 RFLP probes distributed throughout the rice genome, the gene was tagged by a single copy DNA probe, RG4, which was located on chromosome 7. The genetic distance between the salt tolerant gene and RG4 was 7.0 ± 2.9 cM. Based on the split method, a method which could be currently used to evaluate the damage of salt stress in rice was proposed.     

Alcohol and aldehyde dehydrogenase genotypes and drinking behavior of Chinese living in Shanghai

Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), the principal enzymes responsible for oxidative metabolism of ethanol, exist in multiple, genetically determined molecular forms. Widely different kinetic properties in some of these isozymes account for the individual differences in alcohol sensitivity. In this study we used the polymerase chain reaction/restriction fragment length polymorphism method to determine the genotypes of the ADH2 and ALDH2 loci of alcoholic and nonalcoholic Chinese living in Shanghai. We also investigated the subjects' drinking patterns by means of semistructured interviews. The alcoholics had significantly lower frequencies of the ADH2² and ALDH2² alleles than did the nonalcoholics, suggesting the inhibitory effects of these alleles for the development of alcoholism. In the nonalcoholic subjects, ADH2² had little, if any, effect, despite the significant effect of the ALDH2² allele in decreasing the alcohol consumption of the individual. Taken together, these results fit the proposed hypothesis for the development of alcoholism, i.e., drinking behavior is greatly influenced by the individual's gentoypes of alcohol-metabolizing enzymes, and the risk of becoming alcoholic is proportionate with the ethanol consumption of the individual.     

蓟属两种植物的染色体研究

本文对蓟属（Ｃｉｒｓｉｕｍ）的两个形态相似的近缘种大刺儿菜和小刺儿菜进行了染色体研究,其中后者为首次报道。观察结果表明：两个种的染色体数目均为２ｎ＝２ｘ＝３４：它们的核型是：大刺儿菜．２ｎ＝２ｘ＝３４＝２０ｍ＋１２ｓｍ＋２ｓｔ：小刺儿菜．２ｎ＝２ｘ＝３４＝２２ｍ＋１０ｋｍ（２ＳＡＴ）＋２ｓｔ。通过核型比较,认为它们是两个独立的种．而且后者比前者进化。     

大刍草稀有等位基因促进玉米密植高产

密植是提高作物单位面积产量、促进粮食增产的重要途径之一。叶夹角是影响玉米(Zea mays)密植的关键因子。中国农业大学田丰课题组最近克隆了2个调控玉米叶夹角的数量性状位点(QTL)——UPA1和UPA2, 揭示了这2个位点的功能基因(brd1和ZmRAVL1)通过油菜素内酯(BR)信号通路调控叶夹角。UPA2位于ZmRAVL1上游9.5 kb, 可与DRL1蛋白结合。另一个影响玉米叶夹角的蛋白LG1可以激活ZmRAVL1的表达; DRL1蛋白与LG1蛋白直接互作抑制LG1对ZmRAVL1的激活表达。玉米祖先种大刍草(teosinte)的UPA2位点序列与DRL1蛋白结合能力更强, 导致大刍草ZmRAVL1的表达受到更强的抑制, 下调表达的ZmRAVL1进一步使下游基因brd1的表达下调, 进而降低叶环区的内源BR水平, 导致叶夹角变小。将大刍草的UPA2等位基因导入到玉米中或对玉米中ZmRAVL1进行基因编辑, 在密植条件下均可显著提高玉米产量。上述发现为高产玉米品种的分子育种改良提供了重要理论基础和基因资源。     

高效液相层析法纯化人胎盘谷胱甘肽硫转移酶

足月分娩的新鲜胎盘组织制成匀浆后,经高速离心、超速离心,谷胱甘肽(GSH)Sepharose 6B亲合层析,Amicon pM-10膜超过滤及高效液相层析,最终经SDS-PAGE鉴定,结果呈现单一亚基区带,其亚基分子量为25000。 根据我们现有高效液相设备条件,用ODS柱代替RadulovicL等报道的特异阴离子柱,用磷酸盐洗脱液代替含谷胱甘肽、二硫苏糖醇及氯化钾的梯度洗脱液,从人胎盘组织成功地制备了谷胱甘肽硫转移酶(GST)纯酶,全过程在15min内完成,保留时间及主峰面积的重复性均较理想,7次实验结果的变异系数为0.2％,最终纯化578.9倍。本研究为各种形式GST的纯化制备提供了一个新的、重复性好、分辨率高及回收理想的简易方法。     

Chemical modification of Bacillus thuringiensis subsp. thuringiensis (HD-524) trypsin-activated endotoxin: implication of tyrosine residues in lepidopteran cell lysis

A purified protein fraction from a solubilized and trypsin-digested extract of Bacillus thuringiensis subsp. thuringiensis (HD-524) fermentation powder was lytic to cells from several lepidopteran lines. Maximum yield was obtained by alkaline carbonate-thiocyanate solubilization of washed powder followed by trypsin digestion and Sephacryl (S-300) chromatography. The alkaline carbonate-solubilized fraction consisted predominantly of two bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with MW of 144 +/- 0.9 kDa and 134 +/- 1.4 kDa. After trypsin treatment and column chromatography, the cytolytic fraction consisted of a major band with a MW of 60.0 +/- 1.8 kDa and a minor band of 69 +/- 0.9 kDa. Cells from Trichoplusia ni (TN368) were most susceptible to lysis with 50% of cells lysed at 3 micrograms/ml, followed by Spodoptera frugiperda cells (SF21AE) exhibiting 50% cell lysis at 5 micrograms/ml and Lymantria dispar cells (Ld652Y) showing 40% lysis at 10 micrograms/ml. Chemical modification of the polypeptides was performed to determine the role of certain amino acid residues in the cytolytic activity. The group-specific reagent tetranitromethane was used to nitrate and oxidize tyrosine and cysteine residues, respectively. Cysteine residues alone were also modified with p-hydroxymercuribenzoic acid. Lysine residues were modified with O-methylisourea. Of the three types of amino acid residues, only the modification of tyrosine resulted in reduced cell lysis.