Studies inin situ PCR detected by the BrdU antibody technique

Using the BrdU antibody technique followed by an immuno-chemical staining (BAT), the amplification of DNA fragments specific to human Y chromosome on cell specimen slides was efficiently detected. Whether direct BrdU incorporation into PCR products orin situ hybridization with PCR products on slides, the amplified target DNA fragments of specimen were visualized by BAT under the microscope. The availability of BAT and differences in the sensitivity and efficiency between BAT and dig-11-dUTP labeling in cellin situ PCR were discussed. Zhang Xiyuan: born in Oct. 1935, Professor, Current research interest in Cellular and Molecular Biology Supported by the National Natural Science Foundation of China     

In situ hybridization of rDNA on chromosomes of the honeybee,Apis mellifera L.

DNA probes containing the repeated rDNA region ofDrosophila melanogaster (coding for e.g. 28S and 18S rRNA) hybridized in situ to distinct regions of two heterologous mitotic chromosomes of the honeybee, identifying the nucleolus organizing regions (NORs). The method allows a rapid establishment of a physical map ofApis mellifera using other DNA probes ofDrosophila. This is the first report on well-defined chromosomal markers in the honeybee.     

Transfer of bacterial blight resistance from <Emphasis Type="Italic">Oryza meyeriana</Emphasis> to <Emphasis Type="Italic">O. sativa</Emphasis> L. by asymmetric somatic hybridization

Asymmetric somatic hybrid plants were produced between cultivated rice (Oryza sativa L.) and wild species [O. meyeriana (Zoll. etMor, exSteud.)] with high resistance to rice bacterial blight. X-ray-irradiated protoplasts of the wild species were used as donor and chemically fused with iodoacetamide-inactivated protoplasts of rice cv. 02428 to produce hybrids. Seventy-two plants were regenerated from 623 calli based on metabolic complementation. The morphological characters of the plants closely resembled that of the rice. Simple sequence repeats were employed to identify their hybridity. Cytological analysis of root-tips revealed that their chromosome number varied in the range of 27--38. The somatic hybrids were inoculated with strains of Xanthamonas oryzae pv. oryzae at adult growth stage and demonstrated the resistance to bacterial blight introgression from the O. meyeriana.     

荧光原位杂交技术在染色体生物学中的应用

荧光原位杂交技术(Fluorescence in Situ Hybridization，简称为FISH)是20世纪80年代开始发展起来的一种新的定位技术．本文简要介绍了荧光原位杂交技术的创立、发展及其在染色体生物学中的应用．     

穿山龙液泡H+-ATPase c亚基基因片段的克隆

利用抑制消减杂交（SSH）和SMART技术，构建了3年生和1年生穿山龙（Dioscorea nipponica）根组织mRNA群体间正向差减cDNA文库．从34个随机测序的cDNA克隆中，发现了1个编码液泡H^＋-ATPase（V-H^＋-ATPase）c亚基的克隆，并命名为DnvHAc1（Accession No：DN792564）．序列同源性分析表明，其cDNA序列长486bp，3’端具有PolyA结构，其编码的氨基酸序列（115个氨基酸残基）也与多种植物的液泡H^＋-ATPase c亚基（16kDa proteolipids）具有较高同源性，具有3个跨膜疏水结构域，结构域Ⅲ为功能保守的区域，有dicyclohexylcarbodimide（DCCD）结合位点，Glu-92在调节液泡H^＋-ATPase具有重要作用．该研究结果为克隆其全长基因和进一步研究其在薯蓣皂甙次生代谢生物合成中的功能提供了重要的实验基础．     

1株碱杆菌属菌株的分离和鉴定

 从新疆采集的盐土样品中分离到1株革兰氏阳性、严格好氧、产孢、嗜盐细菌菌株YIM 012,对其进行了系统分类学研究.菌株YIM 012为杆菌,生长最适温度为28~45℃,最适pH 7.0~8.0,并能在70~250g/L的盐质量浓度中生长良好.其DNA的G+C的摩尔分数为39%,16S rRNA基因序列分析结果显示,该菌株与碱杆菌属Alkalibacillus salilacus(DSM 16460^T)亲缘关系最近,相似性达到99.6%,但与同属其它2个有效种间相似性低于96%.杂交结果显示其与A. salilacus亲缘关系低于20%,并且在生理生化方面也有差异.综合分析,确定该菌株为碱杆菌属的1个新种,将其命名为嗜盐碱杆菌(Alkalibacillus halophilus).