Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells.

Herpes simplex virus type 1 (HSV-1) plasmid vectors have promise for genetic intervention in the brain, but several problems caused by the helper virus have compromised their utility. To develop a helper virus-free packaging system for these vectors, the DNA cleavage/packaging signals were deleted from a set of cosmids that represents the HSV-1 genome. Following cotransfection into cells, this modified cosmid set supported replication and packaging of vector DNA. However, in the absence of the DNA cleavage/packaging signals, the HSV-1 genome was not packaged, and consequently vector stocks were free of detectable helper virus. In the absence of helper virus, the vectors efficiently infected rat neural cells in culture or in the brain with minimal cytopathic effects. beta-galactosidase-positive cells were observed for at least 1 month in vivo, and vector DNA persisted for this period. This system may facilitate studies on neuronal physiology and potential therapeutic applications.     

利用植物激素调控嫁接形成的初步研究

利用黄瓜（Ｃｕｃｕｍ ｉｓｓａｔｉｖｕｓ）试管苗进行离体茎段自体嫁接,研究ＩＢＡ 和６－ＢＡ 对嫁接形成的影响时发现：进行离体茎段嫁接时,用试管苗茎段可简化嫁接过程,减少污染。嫁接茎段的颜色变化、不定根发生和愈伤组织形成与激素浓度有关。植物激素通过影响砧木和接穗间维管束桥形成的时间和数目调控嫁接组合的发育。在作者的实验中,最佳的激素条件是：在接穗培养基中加ＩＢＡ １．２ ｍ ｇ／Ｌ,在接穗和砧木培养基中加６－ＢＡ ０．３ ｍ ｇ／Ｌ。     

菠菜BADH单抗细胞株及其应用初探

用菠菜甜菜碱醛脱氢酶 ( BADH)免疫巴比西 ( BALB/c)小鼠 ,将其脾细胞与骨髓瘤细胞 SP2 /O-Ag1 4融合 ,在 1 92孔中 ,有约 1 4 %孔生长的杂交瘤细胞 ,用间接酶联免疫方法 ( ELISA)检测表现为阳性。选择其中 2 G3和 2 D10 细胞系 ,用有限稀释法进行克隆化培养 ,约 2 0 %克隆化细胞为强阳性。选择其中 2 G3- H3细胞株注射到 BALB/c小鼠腹腔中诱导腹水 ,腹水的单抗效价为 1∶ 1 0 3。应用 BADH单抗检查了大麦、水稻、高粱、小麦幼苗的叶片和根的粗提物 ,均呈阳性反应 ,表明 BADH除在光合组织中存在外 ,在非光合组织中也可能存在。讨论了非光合组织 BADH的意义     

泡沙参同工酶基因位点的遗传分析

利用聚丙烯酰胺凝胶电泳技术 ,对来自天然群体 (居群 )的泡沙参 (Adenophora potaninii Korsh.)及其人工杂交子代进行了 8种同工酶的电泳检测和谱带遗传分析 ,以确定编码这些酶系统的基因位点和等位基因。选用 4种不同的凝胶缓冲系统 ,对下列不同酶系统进行了酶谱的遗传分析 :天冬氨酸转氨酶 (AAT)、酯酶 (EST)、甲酸脱氢酶 (FDH)、谷氨酸脱氢酶 (GDH)、异柠檬酸脱氢酶 (IDH)、乳酸脱氢酶(LDH)、苹果酸酶 (ME)和超氧化物歧化酶 (SOD)。结果表明 ,这 8种酶系统至少由 1 8个基因位点编码 ,其中 1 2个位点为遗传稳定的等位酶位点 ,是可靠的遗传标记。酶谱的分离式样表明 ,EST为单聚体结构 ,AAT、FDH、IDH、SOD为二聚体结构 ,GDH为六聚体结构。最后对同工酶的器官和发育特异性以及同工酶基因位点的遗传分析进行了讨论     

水稻幼芽细胞生物膜上的赤霉素结合蛋白的结合特性

在水稻 (Oryza sativa)幼芽中存在膜结合的赤霉素结合蛋白 ,其与 GA3 结合的平衡解离常数(Kd)为 6.5× 1 0 -8mol/ L,总浓度为 0 .3 pmol· mg-1 蛋白质。结合蛋白与 GA3 结合活力在 0℃时比 2 5℃时高 1 4 0 %。它与 GA3 结合的最适 p H为 5。 GA3 与此结合蛋白的结合量随反应时间延长而增加 ,1 h达最大值 ,以后又逐渐下降。 IAA、ABA可与 GA3 竞争赤霉素结合蛋白。     

大豆结瘤突变体成分的初步分析及其对硝酸还原酶活性和结瘤的影响

超结瘤大豆（Ｇｌｙｃｉｎｅ ｍ ａｘ （Ｌ．） Ｍｅｒｒ．） ｎｔｓ ３８２ 和不结瘤大豆Ｎｏｄ ４９ 的叶和根组织水提取物经Ｓｅｐｈａｄｅｘ Ｇ２５ 过滤、洗脱,再根据洗脱物对硝酸还原酶（ＮＲ）活性的影响可划分为４ 个组分（ｆｒａｃｔｉｏｎ）样品,即ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ１、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ２、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ３ 和ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ４。其中, ｎｔｓ３８２ Ｆ２ 和Ｆ４ 抑制ＮＲ 活性作用在接种ＵＳＤＡ１１０ 后明显下降, 但接种的ｎｔｓ ３８２ Ｆ２ 却能提高大豆Ｂｒａｇｇ 的结瘤数达一倍, 而接种的ｎｔｓ ３８２ Ｆ３ 和Ｆ４ 的作用不明显。ＮＲ 活性抑制因子不是刺激结瘤的因子, 刺激结瘤的因子主要分布在接种的ｎｔｓ３８２ Ｆ２ 部分中。与这一现象相反, Ｎｏｄ ４９ Ｆ２ 和Ｆ４ 抑制ＮＲ活性的作用在接种后更强, 且也抑制大豆ｎｔｓ ３８２ 的结瘤, 其中Ｎｏｄ ４９ Ｆ４ 抑制结瘤的作用基本不能逆转。抑制结瘤因子主要分布在接过种的Ｎｏｄ ４９ Ｆ４ 部分中     

细胞分裂素生物合成基因在调节一组烟草病理相关蛋白基因表达中的作用

对一组病理相关蛋白基因在烟草 ( N icotiana tabacum cv. Wisconsin 38)中的表达情况进行了研究 ,包括 :碱性几丁质酶、β- 1 ,3-葡萄糖苷酶、渗透蛋白及伸展蛋白。RNA杂交实验表明在正常烟草植株中上述 4个基因具有发育和器官专一性的表达。在含有细胞分裂素生物合成基因的转基因烟草丛生芽中 ,这 4个基因的表达受过量合成的内源细胞分裂素和载体效应的共同调节 ,细胞分裂素降低这些基因的表达 ,而载体效应则促进它们的表达。热激处理也明显降低这 4种基因的表达水平。上述结果表明这些病理相关蛋白基因具有复杂的调控系统     

一种快速提取禽源性大肠埃希氏菌外胰蛋白的方法

介绍一种快速提取禽源性大肠埃希氏菌外膜蛋白的方法。该法是对Ｋａｐｕｒ等发表的方法的改进。全过程只需超速离心一次,比Ｋａｐｕｒ等的方法缩短了４ｈ。所得样品可直接用于禽源性大肠埃希氏菌外膜蛋白模式的测定。     

Optimal conditions for levan formation by an overexpressed recombinant levansucrase

Summary A genetically modified levansucrase, which contained His-affinity tag in its C-terminal, was constructed by PCR reaction using two synthetic primers. This modified protein was produced up to 30 % in total cell protein of E. coli, and was purified by a one-step affinity chromatography. The optimum pH for levan production was pH 5 and the optimum temperature was 0 °C. The higher velocity of levan formation within shorter enzyme reaction times was achieved by increasing the levels of enzyme concentration. The optimal sucrose concentration for levan production was around 20 %. Under these conditions, more than 50 g levan/l was produced.     

Isolation of a root-specific cDNA encoding a ns-LTP-like protein from the roots of bean (Phaseolus vulgaris L.) seedlings

A root-specific cDNA clone, PVR3, was isolated from a bean (Phaseolus vulgaris L.) root cDNA library by a differential screening procedure. The nucleotide sequence of PVR3 contains an open reading frame coding for an 11.14 kDa polypeptide of 102 amino acid residues; the first 25 amino acids correspond to the sequence characteristic of a signal peptide. Comparison of the deduced PVR3 polypeptide sequence with the polypeptide sequences of previously cloned genes indicates that PVR3 may encode a ns-LTP-like protein. Molecular modelling of the PVR3 protein predicts that it has a three-dimensional structure that is similar to the three-dimensional model determined from the maize ns-LTP. The PVR3 mRNA accumulated mainly in the roots of young seedlings. It can be detected at low levels in flowers, but it is not detected in other organs. Genomic Southern blot analysis indicates that the genomic DNA corresponding to PVR3 cDNA is encoded by a single gene or small gene family in the bean genome.