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141.
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143.
Protoplasts isolated from both 7-day-old light-grown and 4-day-old dark/dim light-grown cotyledons of four Brassica campestris varieties (Arlo, Sonja, Bunyip and Wonk Bok) were cultured in three liquid media: modified K8P, modified MS and modified Pelletier's B to compare the capacities for cell division and plant regeneration. Following cell wall regeneration the cultured protoplasts from dark/dim light-grown cotyledons of four varieties showed rapid division and high frequency of cell division compared with those isolated from light-grown cotyledons. The frequencies of cell division were significantly influenced by varieties and culture media but only in cultured protoplasts isolated from dark/dim light-grown cotyledons. The interaction between varieties and media was also significant. Cell colonies formed within 7–14 days in protoplast cultures from dark/dim light-grown cotyledons, and calli subsequently grown on a solid medium developed shoots when transferred onto a regeneration medium. Three of four tested varieties (Arlo, Sonja and Bunyip) showed shoot regeneration within 2–3 months after protoplast isolation, with a high degree of reproducibility in Arlo and Bunyip. Regenerated shoots, which were induced to root on half-strength MS medium with 0.1 mg.l–1 IBA, survived in soil and grew to produce siliques and set viable seeds in the greenhouse. The present report is the first to document the production of regenerated plants that set seeds in Brassica campestris from cotyledonary protoplasts.Abbreviations BAP
benzylaminopurine
- CPW
Composition of Protoplast Washing-solution
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EDTA
ethylenediamine-tetraacetic acid
- GA3
gibberellic acid
- IBA
indole-3-butyric acid
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog medium
- NAA
-naphthaleneacetic acid
- KT
kinetin
- FDA
fluorescein diacetate
- SDS
sodium dodecyl sulfate 相似文献
144.
Zhao Ying Hiromasa Tojo Takanori Komatsubara Manabu Nakagawa Masami Inada Sumio Kawata Yuji Matsuzawa Mitsuhiro Okamoto 《生物化学与生物物理学报:疾病的分子基础》1994,1226(2):201-205
Enzyme activity, protein contents, and mRNA contents of group II phospholipase A2 (PLA2) in hepatocellular carcinoma (HCC) surgically obtained from 8 patients were compared with those in either its neighboring liver tissues or control liver tissues. The PLA2 specific activity towards the mixed micelles of 1-palmitoyl-2-oleoyl-phosphatidylglycerol and cholate was significantly greater in the tumor tissues (6.62 ± 1.46 nmol/min/mg) than those in the surrounding liver tissues (1.33 ± 0.22 nmol/min/mg) and controls (0.43 ± 0.04 nmol/min/mg). The results of immunoblot analysis using a specific anti-human group II PLA2 antibody and of Northern blot analysis using a human group II PLA2 cDNA as a probe demonstrated that group II PLA2 was responsible for the increased enzyme activity. The contents of immunoreactive group II PLA2 in the tumor tissues (8.81 ± 1.24 ng/mg) were significantly higher than those in the surrounding liver tissues (1.77 ± 0.27 ng/mg); those in the control tissues were below the analytical range of the method used. The group II PLA2 mRNA was also significantly increased in the tumor tissues, compared with that in the surrounding liver tissues, whereas it was not detectable in th controls. This indicates that group II PLA2 in HCC is induced at the pretranslational level. 相似文献
145.
马蹄香大小孢子发生及雌雄配子体形成 总被引:1,自引:0,他引:1
马蹄香(Sarumahenryi,Oliv.)花药壁的发育属双子叶型。花粉母细胞减数分裂为同时型,四分体主要为四面体形,少数为左右对称式排列。腺质绒毡层,其细胞可排列为不规则的两层,双核或多核。到单细胞花粉阶段,绒毡层细胞内切向壁上出现许多乌氏体。成熟花粉为2细胞型,圆球状,具单萌发沟。雌蕊6心皮,上部彼此分离、下部联合。倒生胚珠,双珠被,厚球心。胚囊发育蓼型。成熟胚囊为七细胞结构,但两个助细胞退化较早。 相似文献
146.
147.
解离增强镧系元素荧光免疫分析灵敏度的改进 总被引:1,自引:0,他引:1
为提高解离增强镧系荧光免疫分析(DELFIA)的灵敏度或信/噪比,进行了一些重要的方法学研究.观察到了对于不同的铕量,荧光响应和信/噪比都随着增强液体积而明显地变化.对于确定的铕量,存在一个最佳体积,且铕量越小,其最佳体积也越小.实验中选择最佳体积是重要的.研究了增强液制备技术,发展了微滴定板条有效的清洗和干燥方法,使本底荧光明显降低. 相似文献
148.
人重组IL6/IL2融合蛋白的变性、复性及纯化 总被引:1,自引:0,他引:1
经超声破碎,分离已表达CH925包涵体,较系统地研究变性剂浓度、融合蛋白浓度对蛋白折叠的影响.在还原型及氧化型谷胱甘肽复性条件下,成功地将融合蛋白CH925折叠成具有IL6及IL2双活性蛋白,IL6的比活为2.3×108U/mg, IL2比活为2.2×106U/mg.经阴离子交换、凝胶过滤层析,获得一定纯度的CH925,配合反相HPLC.洗脱收集蛋白峰,CH925纯度为98%. 相似文献
149.
鲑鱼生长激素基因分泌型表达质粒的构建 总被引:1,自引:0,他引:1
生长激素(GH)是动物垂体前叶分泌的一种多肽类激素.应用分子重组及PCR等技术,构建了一种鲑鱼生长激素基因分泌型表达质粒pOsGH153,使编码鲑鱼生长激素成熟肽的序列克隆在大肠杆菌分泌型表达载体PIN-Ⅲ-ompA内,直接位于编码大肠杆菌外膜蛋白A信号肽序列的下游,在Lpp-Lac杂合启动子控制下,经IPTG诱导,分子量约23 000的鲑鱼生长激素在大肠杆菌中获得高效表达,该产物具有天然鲑鱼生长激素的免疫活性,直接分泌到细胞周质,而信号肽被自动剪除. 相似文献
150.
豌豆种子发育过程中DNA和RNA的变化 总被引:2,自引:2,他引:0
实验结果表明:豌豆种子发育过程中单粒鲜重、DNA和RNA含量都随开花后日数增加而增加。其中豌豆开花第21天,RNA含量达最高峰,拖尾现象消失而趋于单一带区,DNA的迁移率也降到最低,从而确定出豌豆开花后第21天是基因转录活动降低的转折点,为种子蛋白质基因工程研究提供参考。 相似文献