48例原发性闭经患者的细胞遗传学分析

本文报告对48例原发闭经患者的临床和细胞遣传学分析,共发现染色体异常17例,占35.4%,其中包括45,X,7例;45,X/46,XX,2例;X染色体结构异常5例;核型中有Y染色体3例。讨论了原发闭经的细胞遗传学病因及异常核型与表型的关系。     

细小病毒非结构蛋白转基因小鼠模型建立

将小鼠乳腺癌病毒启动子控制的细小病毒非结构蛋白基因(长5.7kb)氯化铯超速离心,纯化透析后用显微注射法导入C57BL/SJL F_1小鼠受精卵雄核,植入假孕母鼠输卵管,得成活小鼠15只。抽取鼠尾DNA,对其中10只小鼠作PCR和southern blot鉴定,其中4只(40％)整合有目的基因。对首建者B_6()的8只子代小鼠鉴定,3只(37.5％)整合有目的基因。说明导入的目的基因能传代。     

Inheritance of foreign genes in transgenic bean (Phaseolus vulgaris L.) co-transformed via particle bombardment

Exploiting the biolistic process we have generated stable transgenic bean (Phaseolus vulgaris L.) plants with unlinked and linked foreign genes. Co-transformation was conducted using plasmid constructions containing a fusion of the gus and neo genes, which were co-introduced with the methionine-rich 2S albumin gene isolated from the Brazil nut and the antisense sequence of AC1, AC2, AC3 and BC1 genes from the bean golden mosaic geminivirus. The results revealed a co-transformation frequency ranging from 40% to 50% when using unlinked genes and 100% for linked genes. The introduced foreign genes were inherited in a Mendelian fashion in most of the transgenic bean lines. PCR and Southern blot hybridization confirmed the integration of the foreign genes in the plant genome.     

Production and identification of somatic hybrids between Solanum tuberosum and S. papita by using the rolC gene as a morphological selectable marker

A successful hybridization of a diploid clone of Solanum tuberosum with a rolC-transgenic, diploid S. papita clone is reported. By using leaf expiants of this S. papita clone, which after transformation expressed kanamycin resistance, intact protoplasts were obtained, but these protoplasts did not develop to microcalli or regenerate to mature plants. However, protoplasts of the S. tuberosum clone showed a high capacity to regenerate plants from isolated protoplasts. On a medium containing Kanamycin only calli regenerated to plants, which revealed a rolC phenotype (reduced apical dominance with a large number of adventitious shoots and a pale green color of leaves) and later on turned out to be true hybrids. Self fusions of S. papita never developed to microcalli and those of S. tuberosum ceased to develop on the kanamycin-containing medium. Identification of somatic hybrids was done by RFLP and RAPD analysis. In the greenhouse, out of four selected hybrids only FK3.1 was successfully crossed with two standard S. tuberosum varieties (Datura, Desirée). Out of all the seeds germinated, only rolC-negative F¹ seedlings were further characterized. Within the seedling population obvious differences were evident in respect of the S. papita and S. tuberosum characteristics.     

Transformation of Orychophragmus violaceus Using Agrobacterium tumefaciens And Regeneration of Transgenic Plants

ＴｒａｎｓｆｏｒｍａｔｉｏｎｏｆＯｒｙｃｈｏｐｈｒａｇｍｕｓｖｉｏｌａｃｅｕｓＵｓｉｎｇＡｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓＡｎｄＲｅｇｅｎｅｒａｔｉｏｎｏｆＴｒａｎｓｇｅｎｉｃＰｌａｎｔｓａ￥ＺＨＯＵＪｉ－ｍｉｎｇ（周冀明）;ＷＥＩＺｈ...     

Isolation of the facA (acetyl-CoA synthetase) gene of Phycomyces blakesleeanus

A 5.6 kb DNA fragment from the fungus Phycomyces blakesleeanus has been cloned and sequenced. The fragment contains a gene that probably codes for the enzyme acetyl-coenzyme A synthetase (facA). The amino acid sequence deduced for the P. blakesleeanns protein is highly homologous to those of acetyl-coA-synthetases from other organisms. When placed under the control of a constitutive promoter from Aspergillus nidulans, the cloned gene complemented a facA mutation of this organism. In P. blakesleeanns, the expression of facA is induced by acetate.     

Genetic manipulation of 6-phosphofructokinase in potato tubers

The aim of this work was to discover whether genetic manipulation of 6-phosphofructokinase [EC 2.7.1.11; PFK(ATP)] influenced the rate of respiration of tuber tissue of Solanum tuberosum L. Transgenic plants were produced that contained the coding sequence of the Escherichia coli pfkA gene linked to a patatin promoter. Expression of this chimaeric gene in tubers resulted in a 14to 21-fold increase in the maximum catalytic activity of PFK(ATP) without affecting the activities of the other glycolytic enzymes. Tubers, and aged disks of tuber tissue, from transformed plants showed no more than a 30% fall in the content of hexose 6-monophosphates; the other intermediates of glycolysis increased threeto eightfold. Fructose-2,6-bisphosphate was barely detectable in aged disks of transformed tubers. The relative rates of ¹⁴CO₂ production from [1-¹⁴C]-and [6-¹⁴C]-glucose supplied to disks of transformed and control tubers were similar. Oxygen uptake and CO₂ production by aged disks of transformed tubers did not differ significantly from those from control tubers. The same was true of CO₂ production, in air, and in nitrogen, for tuber tissue. It is concluded that PFK(ATP) does not dominate the control of respiration in potato tubers.Abbreviations Fru2,6bisP fructose-2,6-bisphosphate - FW freshweight - GUS -glucuronidase - PFK(ATP) 6-phosphofructokinase - PFK(PPi) pyrophosphate: fructose-6-phosphate 1-phosphotransferase     

Light-stimulated proton transport into the vacuoles of leaf mesophyll cells does not require energization by the tonoplast pyrophosphatase

Photosynthetic characteristics of transgenic tobacco (Nicotiana tabacum L.) plants with a soluble pyrophosphatase in the cytosol of their leaf cells were compared to those of wild-type plants. Although the development of the transgenic plants was somewhat retarded compared to the wild type, as shown by stunted growth and delayed flowering, photosynthetic responses were comparable in transgenic and wild-type leaves of similar physiological age. In particular, light-dependent proton transport into the vacuoles of leaf mesophyll cells was not decreased in leaves of the transgenic plants, which did not contain pyrophosphate in the cytosol owing to the presence of a soluble pyrophosphase. This shows that light-stimulated proton pumping did not require the pumping activity of the tonoplast pyrophosphatase. Apparently, light-stimulated proton pumping can be based solely on the activity of the tonoplast ATPase.Abbreviation CDCF 5-(and 6-)arboxy-2,7-dichlorofluorescein This work was supported within the Sonderforschungsbereiche 176 and 251 of the University of Würzburg.     

根癌农杆菌对甘蓝型油菜的转化及转基因植株的再生

用根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍ ｅｆａｃｉｅｎｓ）共培养法把外源基因导入甘蓝型油菜（Ｂｒａｓｓｉ－ｃａ ｎａｐｕｓＬ．）主要栽培品种“云北２ 号”,获得转基因植株。所用外植体为带有１—２ ｍ ｍ 子叶柄的完整子叶,根癌农杆菌为Ａ２０８ＳＥ（ｐＴｉＴ３７－ＳＥ, ｐＲＯＡ９３）。Ｔｉ质粒ｐＲＯＡ９３ 带有ＮＰＴⅡ及ＧＵＳ嵌合基因。共培养２ 天后转到附加２５ ｍ ｇ／Ｌ卡那霉素的分化培养基（ＭＳ＋ ４．５ ｍ ｇ／ＬＢＡＰ）上。ＡｇＮＯ３ 和羧苄青霉素促进芽的分化,头孢霉素则有抑制作用。最高转化频率为２７％ 。把分化出的茎芽切下,插入含有２５ ｍ ｇ／Ｌ卡那霉素的生根培养基中。羧苄青霉素不利于根的形成。把完整抗性植株移入盛土壤的盆中,生长状况良好。测定β－葡糖苷酸酶活性,８４％ 明显高于对照。以ＮＰＴⅡ基因作探针进行Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分析,证实外源基因已插入到植物细胞基因组中