抗NPV—核酶转基因蚕的研究

将线性化的含核酶基因的重组质粒ｐＧＬ２Ｒｚ用基因枪导入家蚕早期受精卵中,检测了Ｇ１代五龄蚕血淋巴的荧光素酶活性,并且从Ｇ２代开始通过家蚕核多角体病毒（ＮＰＶ）攻击来筛选抗性较高的家蚕进行传代。在Ｇ４代获得一区ＮＰＶ抗性强度比对照组高１０倍的转基因家蚕。     

遗传转化花粉长成单倍体植株

日本Ｈｉｒｏｓｈｉｍａ大学和Ｋａｇａｗａ大学的Ｍ．Ｎｉｓｈｉｈａｒａ和Ｈ．Ｍｏｒｉｋａｗａ及其同事报道,他们从用粒子轰击法转化的花粉第一次获得了稳定的转基因单倍体植株。Ｎｉｓｈｉｈａｒａ等人用含有新霉素转移酶Ⅱ（ＮＰＴⅡ）和β－葡糖苷酸酶（ＧＵＳ）基因的质粒ＤＮＡ包被的金粒,转化烟草属Ｎｉｃｏｔｉａｎａｒｕｓｔｉｃａｎａ的未成熟花粉。从被轰击的花粉再生了卡那霉素抗性花粉胚胎,并从总数为２×１０６个被轰击细胞中获得两个独立的转基因株系。一个同时具有ＮＰＴⅡ和ＧＵＳ活性,另一个只有ＮＰＴⅡ活性。两个…     

大麻哈鱼生长激素基因Ⅱ的分子克隆和序列分析

以λ噬菌体ＥＭＢＬ－３为基因载体,构建大麻哈鱼（Ｏｎｃｏｒｈｙｎｃｈｕｓ　ｋｅｔａ）基因组文库,并从中分离出含全长的生长激素（ｃｓＧＨ）基因的克隆。本研究首次报道了对ｃｓＧＨ全长核苷酸序列分析的结果。发现ｃｓＧＨ基因由６个外显子和５个内含子组成,长度为３３６１碱基对（ｂｐ）。由该序列可推导出含２１０个氨基酸的多肽,其中存在２２个疏水氨基酸残基的信号肽。本研究所分析的ｃｓＧＨ的外显子和内含子,其排列方式与其他鲑类（Ｓａｌｍｏｎｉｄｓ）和罗非鱼（Ｔｉｌａｐｉａ）是相似的,而与鲤科鱼类（Ｃａｒｐｓ）不同,后者外显子和内含子的排列方式与哺乳类和鸟类相似。将所测定的 ｃｓＧＨ基因的编码区与已发表的 ｃｓＧＨⅡ和ｃｓＧＨ Ⅱ两种 ｃＤＮＡ序列进行比较,证明本研究所克隆的 ｃｓＧＨ基因应属于 ｃｓＧＨ Ⅱ,因它们的编码序列 １００％同源。     

紫云英细胞转化条件的研究

研究了紫云英（ＡｓｔｒａｇａｌｕｓｓｉｎｉｃｕｓＬ．）细胞遗传转化的条件。根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍ ｅｆａｃｉｅｎｓ）经含乙酰丁香酮的低ｐＨ／ＰＯ３－４ 诱导培养后,用来感染紫云英下胚轴原生质体,随后的细胞ＧＵＳ瞬间表达活性显著提高,间接证明了上述预培养诱导活化了细菌ｖｉｒ基因,促进了Ｔ－ＤＮＡ 向植物细胞转移。在ＰＥＧ介导的ＤＮＡ 转移中,较高的ｐＨ 和Ｃａ２＋ 浓度能够提高细胞ＧＵＳ活性。质粒ＤＮＡ 浓度及启动子类型对外源基因在植物细胞内表达也有一定影响。采用外植体－农杆菌共培养法,获得ＧＵＳ和ＮＰＴ Ⅱ基因稳定表达的紫云英转化植株     

豌豆5SrRNA的cDNA克隆、全序列及结构特征分析

以赤霉素（ＧＡ）处理后的Ｇ２豌豆为材料,构建了一个２．０×１０６的ｃＤＮＡ文库,用随机筛选法从该ｃＤＮＡ文库中得到一个完整的ｃＤＮＡ．其中１１１５ｂｐ全序列分析表明,它编码了豌豆５Ｓ核糖体ＲＮＡ（５ＳｒｉｂｏｓｏｍａｌＲＮＡ,５ＳｒＲＮＡ）,基因内部存在着与核糖体蛋白Ｌ５及５ＳｒＲＮＡ结合蛋白（５ＳｒＲＮＡＢｉｎｄｉｎｇＰｒｏｔｅｉｎ）同源的区段,这些同源区段是蛋白与ＲＮＡ结合的位点．基因内部还存在着大量的重复序列．     

编码天麻抗真菌蛋白cDNA的分子克隆

用重组ＤＮＡ 技术研究了编码天麻抗真菌蛋白（ＧＡＦＰ）的基因,从天麻（Ｇａｓｔｒｏｄｉａ ｅｌａｔａＢｌ．）块茎中提取Ｐｏｌｙ（Ａ） ｍ ＲＮＡ 后合成ｃＤＮＡ,构建成表达型ｃＤＮＡ 文库,用纯化的蛋白质探针通过免疫筛选找出对应的ｃＤＮＡ 克隆。在进一步证明所选用的ｃＤＮＡ 克隆含有重组的λ－ｐｈａｇｅ ＤＮＡ 后,提取和纯化含有插入片段重组子的ＤＮＡ,用Ｅｃｏ ＲＩ酶切分析可见插入片段。已分离出编码天麻抗真菌蛋白的基因     

雪花莲外源凝集素基因转化番茄

将含有雪花莲外源凝集素基因（ＧＮＡ）的质粒ｐＲＳＳＧＮＡ１通过冻融法转化到根癌土壤杆菌９Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）菌株ＬＢＡ４４０４中。采用叶盘法转化番茄（Ｌｙｃｏｐｅｒｓｉｃｏｎ ｅｓｃｕｌｅｎｔｕｍ Ｍｉｌｌ．）栽培品种“Ｃ８”、“Ａ３９”和“Ａ５３”,获得了含ＧＮＡ基因的４３株转化植物株。通过卡那霉素抗性鉴定、ＮＰＴⅡ基     

大球母贝精子介导外源基因转移研究

将大球母贝（Ｐｉｎｃｔａｄａ ｍａｘｉｍａ Ｊａｍｅｓｏｎ）精子与“全鱼”ＧＨ基因重组体ｐＣＡｇｃＧＨ和ｐＣＡｇｃＧＨｃ的线性ＤＮＡ混合,温育３０ｍｉｎ,经６次、２＾７、１０ｋＶ脉冲电处理后,与卵子受精,得到若干贝苗。从贝苗中提取ＤＮＡ,经ＰＣＲ扩增和Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分子杂交表明,部分受体带有外源基因,当与精子温育的外源基因浓度分别为２μｇ／ｍＬ,６μｇ／ｍＬ及１８μｇ／ｍＬ时,相应贝苗     

含人TNF-α、IL-2及NeoR基因多顺反子逆转录病毒载体的构建及表达

利用脑炎心肌炎病毒和脊髓灰质炎病毒的内核糖体进入位点,连接人ＴＮＦ－α及ＩＬ－２ｃＤＮＡ和选择基因新霉素磷酸转移酶基因（ＮｅｏＲ）,使ＴＮＦ－α、ＩＬ－２及ＮｅｏＲ基因均受控于病毒ＬＴＲ启动子,将这３个基因转录至同一ｍＲＮＡ,从而构建成含人ＴＮＦ－α、ＩＬ－２及ＮｅｏＲ基因多顺反子逆转录病毒载体ｐＧＣＥＮ／ＴＲＩ。在ＬｉｐｏｆｅｃｔＡＭＩＮＥ介导下将其导入包装细胞ＰＡ３１７,Ｇ４１８筛选得单克隆,病毒滴度为５×１０５ＣＦＵ／ｍｌ重组病毒分泌的细胞株,经ＰＣＲ证明外源基因已整合至基因组,Ｎｏｒｔｈｅｒｎ印迹显示出单一转录本。用重组病毒上清感染不同的细胞,Ｇ４１８筛选所获得的抗性克隆能不同程度地表达ＴＮＦ－α及ＩＬ－２。实验结果表明,含ＩＲＥＳ的多顺反子逆转录病毒载体能很好地在同一靶细胞中表达多个外源基因。     

抗除草剂基因和荧光素酶基因在小麦愈伤组织中的稳定表达

用ＰＥＧ介导法将抗除草剂基因和萤火虫荧光素酶基因转入小麦原生质体,在含５０ｍｇ／Ｌ Ｂａｓｔａ的培养基上筛选出有抗性的愈伤组织。ＤＮＡ分子杂交结果表明ｂａｒ基因和ｌｕｃ基因都已整合到小麦细胞基因组中,萤火虫荧光素酶活性测定也检测到ｌｕｃ基因的表达。     

Organolanthanoids. XIII. Bis(pentadeuterocyclopentadienyl)ytterbium(II)

The complex (C₅D₅)₂Yb(dme) (dme = 1,2- dimethoxyethane) has been prepared by reaction of Tl(C₅D₅) with ytterbium metal in dme and is isostructural with (C₅H₅)₂Yb(dme). Thermal desolvation under vacuum yields (C₅D₅)₂Yb, for which essentially identical high resolution neutron powder diffraction data were obtained at 294 K and 77 K, but the structure could not be determined.     

AGRILUS CUPRIPES (CARTER) COMB. N. (COLEOPTERA: BUPRESTIDAE)

i
Synechocera cupripes Carter is transferred to Agrilus Stephens.
Subfamily BUPRESTINAE
Tribe AGRILINI
Genus Agrilus Stephens
Agrilus Stephens, 1830, Ill. Brit Ent. 3: 239.
Agrilus cupripes (Carter) comb.n.
Synechocera cupripes Carter, 1924, Proc. Linn. Soc. N.S.W. 49: 31.     

Lindernia kinmenensis sp. nov. (Scrophulariaceae) from Kinmen (Taiwan)

Lindernia kinmenensis Y. S. Liang, C. H. Chen, & C. L. Tasi sp. nov. from Kinmen (Taiwan) is described. The new species belongs to section Torenioides and is most similar to L. crustacea. It differs from the latter by the following characters: calyx pubescent between the ridges (vs pubescent on ridges), with (2)3(4) bristles on the ridge near the apex of the calyx‐lobes (vs without such bristles), corolla shorter, 6–8 mm long (vs 7–11 mm long) and pale purple (vs purple or blue), posterior theca with obtuse (vs acute) apex. This species is usually found in sandy wetlands. A line drawing, colour photos and SEM micrographs of pollen and seed as aids for identification are provided.     

Rhabditis (Oscheius) pheropsophi n. sp. (Rhabditida: Rhabditidae)

Rhabditis (Oscheius) pheropsophi n. sp., associated with cadavers of the bombardier beetle, Pheropsophus aequinoctialis, is described from material collected in Brazil. Mean body length of the female is 1,217 μm, of the male 872 μm, and of the dauer juvenile 568 μm. The female has six lips with one papilla on each lateral lip and two on each sublateral lip; stoma wall thickened dorsally, metarhabdions with warts, excretory pore near base of esophagus, tail long (c = 8), and phasmids prominent, protruding on scanning electron microscope preparations. The male has 10 pairs of bursal ribs, with the terminal pair considerably smaller than the others; spicules fused distally two-thirds of their length. The new species can be distinguished from other members of the Dolichura-group by its fused spicules.     

Rhabditis (Rhabditis) freitasi sp.n. and Rhabditis (Rhabditis) costai sp.n. (Nematoda-Rhabditidae) isolated from bovine otitis

The author described two new species of Nematoda-Rhabditidae collected from the auditory meatus of catle with ear infection: Rhabditis (Rhabditis) freitasi sp.n. from Formosa county in the state of Goiás and Rhabditis (Rhabditis) costai sp.n. from Sert?ozinho county in the state of S?o Paulo.