甘蓝型油菜秦优10号杂交种纯度鉴定的SSR引物筛选

为了建立一套快速可靠的油菜杂交种纯度的鉴定方法,本文以秦优10号及其亲本、杂种ZZH2为试验材料,对前人开发的2对SSR引物(7号引物, 9号引物)进行了再次筛选.结果显示,7号引物不但能很好地区别出混入杂交种中的亲本,还能将杂交种子的同母异父组合种子从杂交种中分离出来,能够用于鉴别秦优10号杂交种子的真伪;9号引物能区分出杂交种和母本,但不能区分开杂交种和父本.同时,本试验利用人工制成的秦优10号杂交种标准样(纯度为100%)以及7份大田鉴定不同纯度梯度的杂交种子对7号引物鉴定结果的准确性进行了验证,鉴定结果与大田鉴定结果基本一致.本文结果将为鉴定秦优10号杂交种纯度提供更准确的技术资料.     

共表达H5N1流感病毒M1和HA基因的DNA疫苗与腺病毒载体疫苗的小鼠免疫评价

为评价在小鼠体内表达流感病毒M1和HA基因诱导的免疫反应,制备共表达H5N1亚型禽流感病毒 (A/Anhui/1/2005) 全长基质蛋白1 (M1) 基因和血凝素 (HA) 基因的重组DNA疫苗pStar-M1/HA和重组腺病毒载体疫苗Ad-M1/HA,将其按初免-加强程序免疫BALB/c小鼠,共免疫4次,每次间隔14 d。第1、3次用DNA疫苗,第2、4次用重组腺病毒载体疫苗,每次免疫前及末次免疫后14 d采集小鼠血清用于检测体液免疫应答,末次免疫后14 d采集小鼠脾淋巴细胞用于检测细胞免疫应答。血凝     

大熊猫的生境选择特征

基于王朗国家级自然保护区1997—2009年的连续监测数据,利用分布频率法和Bai-ley法,从地形因子、森林群落结构和主食竹3个方面研究了大熊猫的生境选择特征.结果表明:王朗国家级自然保护区的大熊猫对生境具有明显的选择性.在地形上,多选择海拔在2500～3000 m的山体脊部、上部和中部的均匀坡和凸坡,坡向西南,坡度在6°～30°,与水源距离>300 m的环境;森林群落结构上,多选择起源为次生林、针阔混交林,微生境为竹林的生境,乔木平均高度在20～29 m,灌木盖度在0～24%;主食竹多选择平均高度在2～5 m,竹丛盖度>50%,混生,生长状况良好的缺苞箭竹.     

RP-HPLC同时测定喙果绞股蓝中芦丁和槲皮素的含量

目的:建立RP-HPLC同时测定喙果绞股蓝中芦丁和槲皮素含量的方法,并揭示其含量动态变化规律。方法:采用大连依利特SinoChromODS-APC18色谱柱(250 mm×4.6 mm,5μm),流动相:甲醇-0.4%磷酸水溶液梯度洗脱,流速:1 mL/min,检测波长:364 nm,柱温:30℃。结果:芦丁在0.1525～3.8120μg范围内线性关系良好,r=0.9998,平均回收率为98.2%(RSD=2.0%);槲皮素在0.0589～1.4720μg范围内线性关系良好,r=0.9999,平均回收率为97.7%(RSD=2.3%)。含量测定结果表明芦丁和槲皮素的含量具季节性动态变化,芦丁8月份含量高,平均质量分数达6.31 mg/g,槲皮素9月份含量最高,平均质量分数达0.86 mg/g。结论:该方法简单,准确度高,为喙果绞股蓝的质量控制提供实验依据,芦丁和槲皮素含量动态变化规律为其开发利用提供参考。     

江西瑞昌石灰岩山区退耕还林对土壤有机碳的影响

以江西瑞昌石灰岩山区退耕还林5年的林地为对象,研究了不同退耕还林模式(林苗一体化、药用纯林、多树种混交和竹阔混交)对土壤有机碳含量的影响.结果表明:与对照相比,退耕还林地0～20 cm土层的土壤有机碳、微生物生物量碳和可矿化碳含量分别增加24.4%、29%和18.4%,退耕还林显著提高了土壤有机碳含量,增强了土壤碳储能力;0～10 cm土层的土壤有机碳、微生物生物量碳和可矿化碳含量极显著高于10～20 cm土层,且退耕还林地土层间差异大于对照;4种退耕还林模式中,林苗一体化模式对土壤有机碳的影响较大.     

血小板活化因子受体研究进展

血汴板活化因子是通过靶细胞膜上的受体而发挥其作用的,该受体属Ｇ联的受体家族,含３４２个氨基酸,有７个疏水的跨膜片段。其作用机制是通过激活磷酯酰肌醇、钙信使系统及相关蛋白激酶,使某些蛋白质发生磷酸化并产生相应的生物学效应。     

Evolving the <Emphasis Type="SmallCaps">l</Emphasis>-lysine high-producing strain of <Emphasis Type="Italic">Escherichia coli</Emphasis> using a newly developed high-throughput screening method

This study provided a new method which applied a selected l-lysine-inducible promoter for evolving lysine industrial strains of E. coli. According to the intracellular levels of the enhanced green fluorescent protein (EGFP) whose expression was controlled by the promoter, 186 strains were preliminarily selected using fluorescence-activated cell sorting from a 10-million-mutant library generated from a l-lysine high-producing E. coli strain. By subsequent multiple parameter evaluation of the 186 selected strains according to the concentration and the yield of lysine, the productivity per unit of cell in 96-deep-well blocks, two mutants MU-1 and MU-2 were obtained. They produced 136.51 ± 1.55 and 133.2 9 ± 1.42 g/L of lysine, respectively, in 5-L jars. Compared with the lysine concentration and the yield of the original strain, those of strain MU-1 improved by 21.00 and 9.05 %, respectively, and those of strain MU-2 improved by 18.14 and 10.41 %, respectively. The mutant selection and evaluation system newly established in our study should be useful for continuous improvement of the current E. coli strains in the lysine industry.     

Immobilization of nitrilase on bioinspired silica for efficient synthesis of 2-hydroxy-4-(methylthio) butanoic acid from 2-hydroxy-4-(methylthio) butanenitrile

In this paper, a simple and effective method using sodium metasilicate as precursor and amine as additive was first reported to immobilize recombinant nitrilase, for efficient production of 2-hydroxy-4-(methylthio) butanoic acid from 2-hydroxy-4-(methylthio) butanenitrile. High immobilization recovery of enzyme activity (above 90 %) was achieved. The immobilized enzyme displayed better thermal stability, pH stability and shelf life compared to free nitrilase. Moreover, it showed excellent reusability and could be recycled up to 16 batches without significant loss in activity. 200 mM 2-hydroxy-4-(methylthio) butanenitrile was completely converted by the immobilized enzyme within 30 min, and the accumulation amount of 2-hydroxy-4-(methylthio) butanoic acid reached 130 mmol/g of immobilized beads after 16 batches. These encouraging results demonstrated the efficiency of the new technology for nitrilase immobilization, which has great potential in preparation of 2-hydroxy-4-(methylthio) butanoic acid.     

CS3菌毛抗原和霍乱毒素B亚基在人伤寒菌中的表达

通过体内外重组的方法,构建了人伤寒菌流行株Ｔｙ２的△ａｒｏＡ,△ａｒｏＣ和ａｓｄ＾－基因缺失突变体（ＲＳ４１７）作为抗原载体菌：同时,构建包含ａｓｄ基因的表达质粒ｐＹＸ１０２,与ＲＳ４１７一起,构成宿主－载全平衡致死系统,用于在没有抗生素条件选择的情况下,稳定表达克隆在表达质粒上的外源抗原基因,将肠毒素性大肠杆菌的菌毛抗原ＩＩＩ（ＣＳ３）基因和霍乱弧菌毒素Ｂ亚基（ＣＴＢ）基因分别克隆至ｐＹＸ１０２