牛乳素Lfcin B基因的合成及其在酵母菌中的表达研究

[目的]为临床研究和治疗某些病原微生物引起的感染提供依据。[方法]参照LfcinB的氨基酸序列,利用化学合成法合成LfcinB基因并克隆到pPIC9K载体中,转化到酵母菌GS115感受态细胞中,用抗性选择标记G418筛选出高拷贝酵母菌转化子。利用甲醇诱导表达重组LfcinB,经Tricine-SDS-PAGE电泳检测目的蛋白牛乳铁蛋白素的表达情况,同时通过体外抑菌试验检测表达产物的抑菌活性。[结果]提取的酵母菌DNA扩增出1条预期的560bp左右的特异性条带。目的基因已经成功克隆到pPIC9K载体中,并转到酵母菌GS115中而且获得了表达。重组酵母表达上清中牛乳铁蛋白素抗菌肽对沙门氏肠炎杆菌的抗菌活性为42.857IU/ml,表明LfcinB基因的酵母表达载体构建成功。[结论]该研究证明利用化学合成法直接合成编码LfcinB的DNA构建真核表达载体、通过生物发酵工程来制备LfcinB是可行的。     

海域霉菌对植物病原真菌抗性的研究

以玉蜀黍平脐蠕孢菌(Bipolaris maydis)和大豆尖孢镰刀菌(Fusarium oxysporum)为指示菌株,采用平板对峙培养法初步测定渤海海洋霉菌的抑菌活性,再用牛津杯扩散法对初筛获得的活性菌株的发酵液进行抑菌活性测定。结果表明,菌株67对玉蜀黍平脐蠕孢菌和大豆尖孢镰刀菌有很强的抑制作用,菌株32对玉蜀黍平脐蠕孢菌有较好的抑制作用。将菌株67在28℃,180 r/min条件下发酵培养66 h,生物量达到7.054 mg/mL,在54~120 h的发酵时间内,发酵液均具有抑菌活性,发酵96 h时抑菌效果最明显,对F.oxysporum和B.maydis的抑菌圈直径分别为20.3 mm和13.6 mm。同时发现,菌株67发酵液的抑菌活性不仅与发酵时间有关,而且与发酵液的pH值相关,具有明显的抑菌活性的发酵液的pH范围为4.0~5.0。     

高活性干酵母对中华鳖非特异性免疫功能和抗病力的影响

报道了高活性干酵母对中华鳖(Pelodiscus sinensis)幼鳖非特异性免疫功能和抗病力影响的研究结果。试验共设6个组，在各组饵料中高活性干酵母的添加量依次为每千克体重0mg(对照组)、600．0mg(试验Ⅰ组)、800．0nag(试验Ⅱ组)、1000．0mg(试验Ⅲ组)、1200．0mg(试验Ⅳ组)和1400．0mg(试验Ⅴ组)。投喂30d后，从每组中各取10只中华鳖幼鳖采血，测定了中华鳖幼鳖血液中自细胞的吞噬活性、血清中溶菌酶和补体活性；另从每组分别取30只中华鳖幼鳖用注射嗜水气单胞菌(Aeromonas hydrophila)的方法行人工感染，检测其抗病力。研究结果表明，各试验组中华鳖幼鳖血液中自细胞的吞噬活性均显著高于对照组。第Ⅰ和第Ⅱ组中华鳖幼鳖血清中溶菌酶活性与对照组之间没有显著性差异，而试验Ⅲ、Ⅳ组和Ⅴ组中华鳖幼鳖血清中溶菌酶活性均显著高于对照组。第Ⅱ、Ⅲ、Ⅳ和第Ⅴ组中华鳖幼鳖血清中补体活性与对照组之间存在显著差异，而第1组与对照组之间无显著性差异。投喂高活性干酵母能增强中华鳖幼鳖对人工感染A.hydrvphila活菌的抵抗力。研究结果证明在中华鳖幼鳖饵料中添加高活性干酵母的量以每千克体重1200．0mg左右为宜。     

四脊裸胞壳Dh菌株液体培养条件下淀粉酶的产生及活性影响因子

对虫生真菌四脊裸胞壳(Emericella quadriclineata)Dh菌株产淀粉酶的条件及理化特性进行了研究.在孢子浓度为1.5×106个/mL的PD培养液中恒温(25±1)℃振荡(120 r/min)培养3.5 d,菌丝生物量最大达5.1 mg/mL,产淀粉酶量达272.42 U/mg;培养液初始pH4~6最有利淀粉酶的产生,产酶量为1 377.20~1 528.99 U/mg菌丝.淀粉酶活性在50℃,pH 6.0条件下最高.在pH4~7范围内20℃下处理20 min或在pH4~7范围内37℃下处理1 h酶活较为稳定,保持在65%以上.40℃下处理20 min酶活保持90%以上.60℃和70℃下处理20 min,酶活保持在50%以下.在70℃下处理80 min,剩余酶活仅为11.70%.一定浓度的Cu2+和Ba2+有利于酶活提高;同时Mg2+,Fe2+,Ca2+,NH4+对酶活性的抑制作用依次增强.     

生物工程应用于作物育种的试验研究 第2报 外源DNA处理蚕豆的细胞学和同工酶的若干效应

以外源DNA处理蚕豆萌动种子,观察根尖,芽尖分裂细胞的染色体畸变,分析根芽的过氧化物酶同工酶(POD)和脂酯同工酶(EST).结果表明。DNA 处理能引起落后断片、染色体桥等染色体畸变,畸变率的多少与 DNA 的浓度无关。DNA 处理也使过氧化物酶同工酶带数及酶活性有增加的趋势,脂酶同工酶的活性却受到抑制。     

天然竹汁对细菌的抑制作用

从竹子中提取天然竹汁,并应用抑菌圈直径测定方法和最低抑菌浓度测定方法,探讨了竹汁对金黄色葡萄球菌、绿脓杆菌、大肠杆菌、克氏肺炎杆菌、志贺氏痢疾杆菌、流感杆菌、肺炎双球菌、甲型链球菌等致病菌的抑制效果。结果表明,与硫酸链霉素相比,竹汁对致病菌的抑菌效果明显,是一种具有开发前景的用于呼吸道类的保健物质。     

风信子花粉活力与育性

对蓝巨人等7个风信子品种进行了花粉活力的研究及子房、胚珠的发育观察。结果表明不同风信子品种的花粉，在4℃条件下，蓝巨人的花粉活力最高，保持时间最长，达45d，其他品种能保持10～25d。蔗糖对花粉管生长具有明显的促进作用，最适的培养基蔗糖浓度为15％。在7个风信子品种中，蓝巨人的花粉管萌发情况最好。4个风信子品种获得了种子。     

三个山羊品种FSHR基因部分序列的克隆与分析

以波尔山羊、莱芜黑山羊、崂山奶山羊为研穷对象,对其FSHR基因5’端调控区及第一外星子部分序列进行克隆测序。结果表明,波尔山羊与莱芜黑山羊、崂山奶山羊三者序列的同源性为98．13%,相应的突变率为1．87%。三个品种间共有27处发生碱基缺失/插入,且碱基突变区段位于基因的5’端区转录启动调控区,主要集中在-210～—290bp之间,莱芜黑山羊与崂山奶山羊之间仅出现3个碱基的突变。     

Effects of Cd and Pb pollution on soil enzymatic activities and soil microbiota

Based on a representative sampling method and pot experiment with different concentrations of Cd and Pd, the enzymatic activities (urease, phosphatase, catalase, invertase), population of bacteria, fungus and actinomycete in the soil, the Cd and Pd pollution status of soil samples (from the wastewater-irrigated area of Baoding suburb) were appraised. Unitary linear and nonlinear curve-fitting optimization models were applied in the research, and the relationship between Pb and Cd causing pollution and enzymatic activities of the tested soils were discussed. The research may provide a theoretical basis for protecting the environment in the region of Baiyangdian Lake, Hebei province, prevent soil pollution, and ascertain biochemical indexes, which reflect soil heavy metal pollution levels. The research results indicated that: (1) there was obvious accumulation of Pb and Cd in the wastewater-irrigated area, also the accumulation in wastewater-irrigated soil is more than that in fresh water-irrigated soil, and accumulation on surface layer was more than that in the lower layer. Pb and Cd contents in the tested soils exceeded the standards of soil background values for some major cities at home and abroad and the world soil Cd and Pb contents range. This means that the tested soil had reached a lightly polluted level; (2) there existed an obvious negative correlation between soil enzymatic activities and Pb and Cd contents in wastewater-irrigated soil, where the soil urease and catalase activities decreased obviously with the increase of Pb and Cd contents in soil. Therefore, the urease and catalase can be considered as biochemical indexes that reflect the degree of soil Pb and Cd pollution; (3) the pot experiments indicated that the influence of Cd on soil enzymatic activities was greater than that of Pb. Generally, the effect of Cd on soil phosphatase, urease, catalase is more obvious than that on invertase, while Pb has a more obvious effect on invertase than Cd; (4) pot experiments of triple cropping showed that, Cd and Pb had an obvious inhibiting effect on microbes. The influence sensitivity of Cd and Pb on different microbes was ordered as bacteria > actinomycete > fungus. Cd had a greater inhibiting effect on microbes than Pb, the influence of Cd and Pb on actinomycete and fungus was relatively smaller, therefore, bacteria can be chosen as an important index that reflects the effect of Cd and Pb on microbe sensitivity.     

嗜线虫致病杆菌Xenorhabdus ne matophilus HB310对3种十字花科蔬菜害虫杀虫活性的研究

嗜线虫致病杆菌XenorhabdusnematophilusHB310是与昆虫病原线虫SteinernemacarpocapsaeHB310共生的细菌。对X.nematophilusHB310杀虫活性的测定结果表明:该共生菌菌液对小菜蛾、菜粉蝶和云斑粉蝶等幼虫均有较高的胃毒杀虫活性,饲喂蘸有该菌液的甘蓝叶片72h后,小菜蛾2龄幼虫、云斑粉蝶1龄幼虫校正死亡率达100%,96h菜粉蝶1龄幼虫、云斑粉蝶4龄幼虫的校正死亡率也分别达到100%。云斑粉蝶1龄幼虫最敏感(LC50为6 4998×105cells/mL),其次为小菜蛾2龄幼虫(LC50为1 3614×106cells/mL)和菜粉蝶1龄幼虫(LC50为2 7418×106cells/mL)。深入研究发现胃毒杀虫活性物质在该菌对数生长初期就开始产生,对数生长中期已达到最大量。生测结果表明:胞内、胞外分泌物中都含有较强的胃毒杀虫活性物质,5 994×108cells/mL的原菌液、细胞及同样浓度的菌液上清和胞内上清120h对1龄菜粉蝶幼虫的胃毒杀虫活性均在90%以上,上清的胃毒活性最低,与原菌液、细胞及胞内上清的胃毒活性差异显著。X.nematophilusHB310的Ⅱ型菌菌液的胃毒杀虫活性较低,只有Ⅰ型菌的1/3左右。