Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

NO对大鼠中脑腹侧被盖区DA神经元的调节

实验观察了微量注射 NOS抑制剂 L -NAME及微量联合注射 NO的前体 L-精氨酸( L-Arg) L-NAME于大鼠中脑腹侧被盖区( VTA)对该部位多巴胺神经元的调节。发现VTA注射 L -NAME( 1 mg/ 5μL )后 ,伏隔核( Acb)多巴胺 ( DA )代谢产物—双羟苯乙酸( DOPAC)水平升高到注射前的 1 2 2 .5% ( P <0 .0 0 1 ) ,小剂量注射 L-NAME( 0 .2 mg/ 5μL)对伏隔核 DOPAC水平无明显影响 ;同样方法联合注射 L-Arg( 3 0 0 μg/ 5μL) L-NAME( 1 mg/5μL)后 ,伏隔核 DOPAC水平无明显变化。结论 :VTA微量注射 L-NAME兴奋了该部位的DA神经元 ,而 L -Arg L -NAME联合注射 ,却不能影响 DA神经元的活动 ,说明 NO可以通过L-Arg-NOS-NO途径参与 VTA多巴胺神经元的调节     

茶多酚(TP)对家禽免疫功能的研究

选用14日龄健康的海佩科内用仔鸡48羽,随机分成试验(24羽)和对照(24羽)两组.试验组在饲粮中添加0.25％的茶多酚(TP),研究TP对鸡免疫功能的作用.试验结果表明,TP对自然感染法氏囊病鸡,有明显提高(p<0.05)鸡血液中红细胞C_(3b)受体和免疫复合物的含量及保护鸡脾脏正常的免疫功能,同时也发现TP对公母鸡法氏囊的重量有不同影响.本试验表明,TP具有提高家禽免疫功能的作用.     

黄皮的生物学特性研究

通过本试验观察表明:黄皮[Clausena Lansium (Lour.)Skeels]的开花与温度有密切关系,温度高,开花过程短;温度低,开花过程长。花穗中的花轴增长速度也随温度升高而加快,花轴增长快,则花轴长,花穗大;反之,花轴增长速度慢,花轴短,花穗小。黄皮开花的顺序是从花穗基部第1—4个分枝开始,每个分枝则先从基部1—8个二次分枝首先开放。31℃是黄皮花粉萌发最适宜温度。在培养基中加入15—20％蔗糖或2.4—D,三十烷醇等生长调节物质,明显可提高黄皮花粉的萌发力。本试验对黄皮的物候期及植物学特性同时进行了研究。     

菜用加工兼用栗永荆3号选育

一般认为我国南方栗仅宜于菜用，加工极易开瓣和糊化，经过几年的筛选和加工试剂，永荆3号品种可生产B－E级标准的糖水栗子罐头。     

辣椒夏季品比试验报告

为了考察我公司新推广品种及新配组合在高温伏旱状况下的适应性，我们于2003年5月7日在公司试验基地进行了品种对比试验。结果表明新推广品种中有九个品种或组合为极耐热或耐热，一个品种耐热性稍差。新配组合中部分表现优良，有推广前景。     

管道3PE防腐涂层补口技术标准及规范对比分析

通过对比国内外热收缩带(套)补口技术规范及标准,分析了各项性能指标间的差异,包括材料性能指标、安装工艺及质量检验.指出管道表面处理及底漆涂装是影响热收缩带(套)安装系统性能的重要因素,提出了工程技术规范编制建议.     

真核表达NDV F基因重组质粒的构建及其在CEF细胞中的转染

通过PCR方法自含NDVZF基因的克隆质粒中扩增NDVF基因 ,将其与真核表达载体pcDNA3..1/V5_His_TOPO重组。经核酸内切酶酶切 ,阳性克隆鉴定准确无误 ,核酸序列测定结果与原始基因比较 ,同源性大于 99% ,启始密码和终止密码未出现变异 ,将该重组质粒命名为pcDNANDVZF。将pcDNANDVZF在脂质体作用下转染CEF细胞 ,用间接免疫荧光试验检测 ,结果证明pcDNANDVZF可在CEF细胞中大量表达F蛋白。