一种改良的转基因甘蔗基因组DNA提取方法

用改良的转基因甘蔗基因组DNA提取方法可从少量的转基因甘蔗叶片中简便快速地提取高质量的DNA,有效地去除甘蔗叶片中的多糖、多酚类和RNA等物质。经核酸蛋白测定仪及凝胶电泳分析表明,该改良方法提取的DNA具有典型的DNA分子标准紫外吸收光谱特点,其A260/A280为1．7-1．9,A260/A230为1．8-2．0,叶片的DNA产量为45-60μg(100mg)^-1,适用于对转基因甘蔗进行PCR、酶切和Southern杂交检测分析。     

担子菌草菇总RNA的快速抽提方法

针对高等真菌草菇(Volvariella volvacea),提供一种改进的简易抽提总RNA的方法。纯化的RNA可直接作如下的各种应用,如RT-PCR、poly A-RNA的富集、差异显示、Northem杂交、引物延伸、eDNA合成、基因表达谱芯片和基因表达谱芯片分析。我们用此方法成功地从草菇中抽提到总RNA,A260,A。为1．7—2．0,A260/A230大于2．0,数值显示RNA的纯度是足够高的(RNA没有被蛋白或苯酚等污染)。电泳图上28S：18S比例约为2：1,显示RNA没有被降解。     

丹参DNA提取方法比较

为从富含多糖、酚类物质的丹参幼叶中提取高质量的核总DNA,比较了4种DNA提取方法对丹参叶片的提取效果,结果表明：改良CTAB法提取的DNA溶液颜色为无色透明、A260／A230为2．137、A260／A280为1．816值最好,是提取丹参基因组DNA的有效方法。     

绢丝丽蚌年龄与生长的研究

绢丝丽蚌一年生长一个生长轮。年轮可肉眼观察贝壳外表面凹陷的生长轮来鉴定,用纵剖贝壳明暗相间层数与打磨后观察棱柱层和珍珠层上的生长轮来验证。绢丝丽蚌10龄以前生长较快,10龄以后生长逐渐减慢。10龄以前年龄(A)与壳长(L)呈直线相关,年龄与壳重(Ws)、体重(W)均呈幂函数相关,其10龄以前的方程式分别为：L=0．8980A 0．8600(r=0．9883),Ws=1．0175A^2.3399(r=0．9997),W=1．3188A^2.3333=0．9997)。10龄以后年龄与壳长、壳重和体重均呈直线相关,其回归方程式分别为：L=0．1817A 7．9085(r=0．9813),Ws=10．7720A 61．1930(r=0．9902),W=13．6960A 78．8690(r=0．9903)。壳长与壳重、体重之间均呈幂函数相关,其相关方程式分别为：Ws=0．6303L^2.4846(r=0．9999),W=0．8181L^2.4775(r=0．9999)。壳重与体重之间呈线性相关,其回归方程式为：W=0．3560 1．2744Ws(r=0．9999)。     

草菇病毒——一种新的食用菌ds-RNA病毒

自草菇(Folvariella~olvacea)子实体中分离到一种等轴对称直径为35rim左右的病毒颗粒。病毒样品经琼脂糖电泳显示一条区带,并具有典型核蛋白的紫外吸收光谱。最大吸收为E257,最低吸收为E230,A260／280=1．96。经SDS-不连续聚丙烯酰胺凝胶电泳解离病毒,出现一条主要衣壳多肽的电泳带,分子量为60,000道尔顿。病毒核酸的最大吸收在258nm,最低吸收在232nn,A260/280=2．0,琼脂糖电泳为一个组分。     

大肠杆菌噬菌体C2的一些物理化学特性

大肠杆菌噬菌体C2是含RNA的微球形噬菌体,直径约22毫微米。提纯的噬菌体在SSC缓冲液中沉降系数为79 S,在260毫微米比吸收值为7．8／毫克／毫升。纸层析分析表明其核酸碱基组成的克分子百分数为：A22．0、U 27．0、G 26．1和C 24．8。其解链温度为5 7．5℃。聚丙烯酰胺凝胶电泳测得RNA分子量为11×106。SDS-聚丙烯酰胺凝胶电泳测得外壳蛋白和A蛋白的分子量分别是13,200和42,000。     

两种适用于葡萄不同组织RNA提取方法的比较

以不同葡萄组织为材料对目前常用的2种总RNA提取方法一改良SDS法和CTAB-LiCl法进行研究。2种RNA提取方法中均不使用酚。采用这两种方法从葡萄不同组织中均成功地提取到RNA,琼脂糖凝胶电泳结果显示28s和18SrRNA条带完整清晰。检测A260/A280 值分布在1．7-2．0之间,A260/A230值分布于1．9-2．3之间,说明RNA质量较高。管家基Actin和ACS5基因的检测表明2种方法所得RNA能够满足RT-PCR和基因克隆等研究需要。改良SDS法的RNA得率是CTAB-LiCl法的RNA得率的2～3倍,而CTAB-LiCl法获得的RNA纯度高。可以根据原料的数量和对RNA质量的要求来选取最佳提取方法。     

大豆低聚糖对肠道双歧杆菌和肠杆菌的促生长作用

目的:观察大豆低聚糖和所含糖对肠道菌群中双歧杆菌、肠杆菌体外促生长作用。方法:按1％大豆低聚糖、0．36％水苏糖、0．52％蔗糖、0．11％棉子糖含量配制培养液,分别接种各试验菌株,在0、12、24小时测定各培养液吸光度A值(分光光度计．550nm)和pH值。结果:经24小时培养,加大豆低聚糖的双歧杆菌标准株和临床分离株培养液A值分别为>1．5和1．307,大于肠杆菌标准株和临床分离株的1．1和1．173,而其pH值小于肠杆菌;加水苏糖的双歧杆菌培养液A值为1．47,大于蔗糖的1．4和棉子糖的0．53。结论:大豆低聚糖促双歧杆菌生长作用大于促肠杆菌生长,水苏糖是促双歧杆菌增殖生长的主要成分。     

小梁切除术中应用丝裂霉素C对角膜内皮细胞的影响

目的：观察小梁切除术中应用丝裂霉素C（MMC）对角膜内皮细胞的影响。方法：收集2010年9月2011年5月在我院行小梁切除术的青光眼患者60例（78眼）,随机分为术中应用丝裂霉素c的36例（46眼）患者为A组,术中不用丝裂霉素c的24例（32眼）为B组。分别观察术前、术后1个月和术后3个月两组眼压（10P）、角膜内皮细胞的密度（co）、平均细胞面积（AVG）及细胞面积变异系数（cv）,分析其数量的改变及两组间的差异。结果：A组术前眼压为（35．4±13．7）mmHg,B组术前眼压为（32．5±13．5）mmHg差异无统计学意义（P〉0．05）,A组术后1个月及术后3个月眼压分别为（15．7±3．7）mmHg、（17．0±3．2）mmHg,均低于B组的（19．4±3．7）mmHg、（20．2±2．1）mmHg,差异有统计学意义（P〈0．05）。A组术前、术后1个月及术后3个月角膜内皮细胞密度分别为（2475±484）个／mm2、（2199±373）个／mm2、（2164±332）个／mm2;平均细胞面积分别为（431．4±67．6）μm2、（480．6±66．8）μm2、（463．8±46．2）μm2;细胞面积变异系数分别为（31．1±7．4）％、（34．4±6．3）％、（31．2±7．5）％;术后1个月及术后3个月各参数与术前比较,差异均有统计学意义（P〈0．05）。B组术前、术后1个月及术后3个月角膜内皮细胞密度分别为（2342±94）个／mm2、（2185＋215）个／mm2、（2074218）个／mm2;平均细胞面积分别为（453．9土94．8）μm2、（516.3±100．8）μm2、（499．81＋106．4）μm2;细胞面积变异系数分别为（30．2土3．0）％、（32．7±2．9）％、（31．4±4．3）％;除术后3个月角膜内皮细胞与术前比较有意义（P〈0．05）外,余参数术后1个月及术后3个月与术前比较差异均无统计学意义（P〉0．05）。术后1个月A组的角膜内皮细胞丢失率为10．4％高于B组的6．1％,差异有统计学意义（P〈0．05）;术后3个月A组的角膜内皮细胞丢失率为11．1％高于B组的10．0％,差异无统计学意义（P〉0．05）。结论：小梁切除术中用丝裂霉素C的降压效果比不用丝裂霉素C的效果好,但短期内前者角膜内皮细胞的丢失率高于后者。     

血管紧张素Ⅱ 1型受体基因A1166C多态性与新疆维吾尔族高血压相关性研究

目的:探讨新疆维吾尔族人群血管紧张素Ⅱ1型受体基因(AT1R)A1166C多态性与原发性高血压之间的关系,了解该基因多态在维吾尔族群体中的分布情况。方法:选择新疆维吾尔族原发性高血压病患者126例,正常血压者143例,应用多聚酶链反应、限制性片段长度多态性技术(PCR-RFLP)时入选样本进行基因分型。结果:AT1R基因A1166C多态符合Hardy-Weinberg平衡;AA、AC各基因型频率在维吾尔族病例组和对照组分别为73%、27%和74．8%、25．2%,差异无统计学意义(P＞0．05),C等位基因频率分别为13．5%和12．6%,差异亦无统计学意义(P＞0．05)。结论:AT1R基因A1166C多态可能不是新疆维吾尔族原发性高血压病的遗传易感指标。     

The Impact of Artisanal Fishery on a Tropical Intertidal Benthic Fish Community

We examined the benthic fishes and artisanal fishery in the intertidal flats of Inhaca Island, Mozambique. Results of a questionnaire indicated that catches had decreased, and that piscivorous fish have disappeared. Results of a catch sampling study indicated that current catch rates are low, < 2 kg person^–1 fishing trip^–1. Use of fishing gear was significantly related to season, diel and lunar tidal phase, and habitat. Forty-eight fish species were observed in the catches with eight species comprising 80% of the catch of 1814 specimens. The annual catch was estimated at 26.2t for the whole bay. Highest fishing pressure was observed in the central section of the bay. A demersal fish survey was carried out with a 2-m beam trawl to sample the fish community. Two different areas were sampled, one area with a low, and one with a high fishing pressure. A total of 19889 fishes were caught comprising 93 species. Gobies dominated the catches and accounted for 56% of all specimens. Fishes were small with a mean standard length of 29mm. The Saco area exhibited the highest catch rates and biomass (maximum of 1040 fish 1000 m^–2 and 1490g 1000 m^–2), and the highest species richness and evenness values. Catch composition was different between the two sampling areas, and was strongly affected by season, but less by habitat. Total fish biomass was estimated at 5.6t for the whole area. Stomach content varied with habitat, and season, and was dominated by benthic invertebrates. The largest estimates of consumption were obtained in the tidal channel and the Zostera beds. Mean consumption of benthic organisms was 1.3g AFDW m^–2 yr^–1. The area seemed to be overfished. The heavily fished areas exhibited lower catch rates, lower proportion of piscivorous fish, increased proportion of small fish, and a decrease in species diversity.     

酿酒酵母菌核糖体RNA沉降系数的初步研究

为研究酿酒酵母菌核糖体RNA(rRNA)的沉降系数,用酶解法和液氮研磨法裂解酿酒酵母菌的细胞壁,Trizol Reagent提取其总RNA,同时提取小白鼠和斑马鱼的总RNA进行比较.经紫外分光光度计检测和甲醛琼脂糖变性胶电泳后,RNA纯度好,条带清晰,无弥散或降解现象.试验发现,与酶解法相比,用液氮研磨法破碎酿酒酵母菌细胞壁提取总RNA所用的成本低,时间少,产率和纯度高,适用于少量样品RNA的提取.同时,酿酒酵母菌与斑马鱼和小白鼠总RNA电泳图谱表明,三者的"18S rRNA"在条带大小方面差异较小,而"28S rRNA"差异较大.利用分析型离心机测得的酿酒酵母菌两个较大rRNA的沉降系数分别为24.7S和18.1S.研究结果表明了真核生物rRNA种类的多样性.     

一种有效的花瓣总RNA的提取方法

利用CTAB法以富含花青素类物质的紫蓝色花瓣为材料提取总RNA,经紫外光谱分析A260/A280比值为1.9~2.0,A260/A230比值约为2.0;电泳检测到了28S、18S和5S rRNA清晰的条带;通过RT-PCR扩增出了目的基因的cDNA片段,说明分离的总RNA能去除色素干扰,纯度和反转录活性较高符合RNA相关实验的要求,是一种经济、有效的花瓣总RNA的提取方法。     

An optimized preparation method to obtain high-quality RNA from dry sunflower seeds

In an attempt to isolate high-quality, intact total RNA from sunflower (Helianthus annuus) seeds for investigation of the molecular mechanisms of mutations, we tested various procedures, using kits, including RNAiso Plus, RNAiso Plus+RNAiso-mate for Plant Tissue, Trizol, and the Qi method, but no high-quality total RNA of high integrity was obtained with any of these methods, probably due to the high content of polyphenols, polysaccharides, and secondary metabolites in mature sunflower seeds. Modifications were made to the Qi method. To avoid polyphenol oxidation, frozen dry seeds free of the seedcase were ground in a mortar with an equal amount of PVP30, and the fine ground powder was transferred to an extraction buffer with 2% PVP30 (w/v), 5% β-mercaptoethanol (v/v) and LiCl (8 M). A sample homogenate was extracted with chloroform prior to acidic phenol-chloroform extraction. The total RNA was precipitated with 1/4 volume of NaAc and 2 volumes of absolute ethanol to prevent contamination by polysaccharides. The yield of total RNA was 29.95 μg/100 mg husked dry seeds; the ratios of A260/A230 and A260/A280 were 2.44 and 2.09, respectively. Electrophoretic analysis clearly showed 28S and 18S ribosomal RNA bands. Using the extracted RNA, a fragment of the actin gene was successfully expressed by RT-PCR. This modified protocol is suitable for isolating high-quality total RNA from sunflower seeds for molecular research.     

黄檗(Phellodendron amuranse)叶片总RNA提取方法研究

以黄檗(Phellodendron amuranseRupr.)叶片为材料,分别利用改进的盐酸胍法、Trizol法、CTAB法提取黄檗叶片总RNA,通过RNA产率、纯度、电泳图谱等分析,确立了1种从黄檗叶片中快速分离总RNA的方法。研究结果表明,改进盐酸胍法所提取的总RNA的A260/A280为1.928,28S和18S条带清晰谱图完整性好,而且具有产率高、时间短、成本低的特点,所提取的总RNA适用于mRNA分离、cDNA文库的建立、Northern杂交等分析。     

Rapid and Reliable Method of Extracting DNA and RNA from Sweetpotato, Ipomoea batatas (L). Lam

A quick, simple and reliable method of extracting DNA from sweetpotato (Ipomoea batatas (L.) Lam.) has been developed. The method was applied successfully for extraction of total DNA from leaves and total RNA from leaves and various tissues. The yield of DNA extracted by this procedure was high (about 1 mg/g leaf tissue). The extracted DNA was completely digested by restriction endonucleases indicating the absence of common contaminating compounds. The absorbancy ratios of A260/A230 and A260/A280 of isolated RNA were approx. 2 and the yield was about 0.2 mg/g fresh wt. CIPK and tublin genes were successfully amplified by RT-PCR, suggesting the integrity of isolated RNA. The total DNA and RNA isolated by this method was of sufficient quality for subsequent molecular analysis.     

提取航天诱变向日葵种子总RNA的一种有效方法

目的:从航天诱变向日葵种子中提取高质量的总RNA.方法:采用改进的SDS法,提取缓冲液与氯仿同时作用液氮研磨材料后,用酸酚-氯仿抽提一次,经LiCl过夜沉淀、DNase I处理、1/2体积的无水乙醇沉淀多糖,最后加入1/10体积的醋酸钠和2倍体积的无水乙醇沉淀总RNA,用琼脂糖凝胶电泳与紫外分光光度法测定产量与纯度,用...