兼具SOD和GPX活力的双功能酶的制备及性质研究

用苯甲基磺酰氟（ＰＭＳＦ）和Ｈ_２Ｓｅ相继处理铜锌超氧化物岐化酶（Ｃｕ,Ｚｎ－ＳＯＤ）,将酶分子中的丝氨酸（Ｓｅｒ）转化为硒代半胱氨酸（ＳｅＣｙｓ）,从而引入了谷胱甘肽过氧化物酶（ＧＰＸ）的催化基团,使其在ＳＯＤ酶活性大部分保留的情况下,具有ＧＰＸ活性,其ＧＰＸ活力是ＰＺ５１活力的３０倍。研究了双功能酶的最佳制备条件,包括ＰＭＳＦ的剂量、反应最适温度及Ｈ_２Ｓｅ处理时间等,并用电子能谱、ＤＴＮＢ等方法测定了双功能酶的硒含量;测定了双功能酶对不同底物的米氏常数及双功能酶的荧光光谱、紫外吸收光谱及稳定性。     

Factors affecting gene delivery by particle bombardment ofDendrobium orchids

Summary Five parameters were examined for their effect on transformation ofDendrobium tissues by microprojectile bombardment. The superpromoter in pBI426 produced at least 1.5 times as many transient transformants as the single cauliflower mosaic virus 35S promoter in pBI121 (37 to 69% vs. 0 to 44%) with dark and frequent GUS (β-glucuronidase) staining. Tissue, genotype, and type of microparticle significantly affected transient GUS activity. Higher expression was seen in protocormlike bodies and in hybrid UH44 compared to etiolated shoots and protocorms and to hybrids M61 and K1329-39. Microparticles of 1.6-μm Bio-Rad gold were more effective than 1.0-μm ASI gold. Transient GUS activity did not differ among protocormlike bodies bombarded using helium propellant pressures of 650, 900, or 1100 psi. Transgenic plants were recovered fromDendrobium UH800 protocormlike bodies bombarded with pBI426-coated, 1.1-μm tungsten particles using an early-model gunpowder-driven apparatus with an estimated stable transformation rate of 11.7%. One transgenic plant ofDendrobium UH44 was recovered from etiolated shoot explants bombarded with pBI121-coated, 1.1-μm tungsten particles using the Dupont PDS-1000 with a stable transformation rate of 0.17%. Positive selection results showed 100 to 200 mg·liter⁻¹ kanamycin to be appropriate for regeneration of transgenic plants from protocormlike bodies, protocorms, and etiolated shoot explants over a 3- to 9.5-mo. period.     

Extractive fermentation of lactic acid by immobilizedLactobacillus casei using ion—exchange resin

Summary A novel method of lactic acid fermentation byLactobacillus casei immobilized in Ca—alginate gels is described, in which an ion—exchange resin packed column is attached to a fermentor for separation of lactic acid from fermentative broth. The technique successfully alleviated the restriction imposed by lactic acid on bacterial growth and product formation. As compared to the conventional batch fermentation, the new fermentation technique enhanced the lactic acid productivity and sugar conversion rate from 0.328g/L·h and 88. 2% to 0.482g/L·h and 98.6%, respectively.     

基因工程链激酶(r—SK)纯化及其单克隆抗体制备和鉴定

采用Rotofor等电聚焦和分子筛技术纯化大肠杆菌表达的重组链激酶(r—SK),其纯度为97%。比活性1×10⁶IU／mg,回收率41％。分析所纯化的r—SK　N端氨基酸顺序证实其1—15个氨基酸顺序与SK基因的核苷酸顺序所示3联密码子一致。以纯化r—SK为抗原,通过杂交瘤技术构建了分泌抗r—SK单克隆抗体(McAb)杂交瘤细胞(4D11),该McAb属IgG1亚型．特异地作用于r-SK和C组口溶血性链球菌分泌的SK。用底物显色法测定该McAb可抑制SK对纤溶酶原的激活。Western blot法证实该McAb能识别分子量为16 250Da的r—SK的CNBr裂解片段。推测SK蛋白中第71残基至第237残基间的氨基酸顺序参与SK与纤溶酶原的结合。     

抗人PAI—1单克隆抗体制备,鉴定和应用

以重组PAI-(rPAI-1)为抗原,通过杂交瘤技术获得6株阳性杂交瘤细胞(AP1、AP2、AP3、AP4、AP5和AP6),并用SPA亲和层析纯化了抗PAI-1单克隆抗体(McAb)。所有McAb均能识别rPAI和天然PAI-1,腹水滴度均为10~6以上。6种McAb对PAI-1亲和常数在3.45×10~7—1.05×10~9mol/L之间。AP2、AP3 McAb能完全抑制PAI-1活性,AP4、AP和AP6只能部分抑制PAI-1活性,AP1则不抑制PAI-1活性。6种McAb中只有AP1、AP4和AP5能识别PAI-1/t-PA复合物。利用AP1、AP3、和AP4 McAb制备免疫亲和柱一步纯化HepG_2细胞分泌的PAI-1,纯度大于98％,回收率92％,纯化倍数51倍。利用抗PAI-1 McAb建立了夹心法ELISA,测定了人血浆PAI-1水平,正常人血浆PAI-1平均含量(±D)为24.7±7.75ng/ml。     

碳源对固定化细胞生产柠檬酸的影响

柠檬酸是利用微生物代谢生产的一种极为重要的有机酸．广泛应用于食品、饮料、化工、冶金、印染等各个领域。在国外,近10年来,利用固定化细胞生产柠檬酸已获得较广泛的研究^〔1－6〕,国内也有学者指出,柠檬酸发酵的趋向是利用固定化细胞进行连续化生产⑺。而国内这方面的研究报道很少〔8,9〕。我们利用海藻酸钙凝胶包埋固定化黑曲霉细胞生产柠檬酸．探讨了碳源种类及其浓度对固定化细胞生产柠檬酸的影响。现将结果报道如下。     

甘薯象空间格局及抽样技术研究

通过空间分布型指数分析,甘薯象对薯块、著株危害空间分布型为随机分布或均匀分布;同时确定了理论抽样数     

褐飞虱五龄若虫翅芽长度与成虫翅型关系的研究

褐飞虱饲养于不同水稻品种、不同生育期以及不同种群密度条件下,5龄若虫翅芽长度与成虫翅型具有稳定的相关性：翅芽长于1．10mm羽化后为长翅型雌虫,介于0．96～1．09mm之间为长翅型雄虫或短翅型雌虫,小于0．95mm为短翅型雄虫。根据5龄若虫翅芽长度结合外生殖器特征,编制了成虫翅型检索表。由此,可在5龄若虫期预测成虫的翅型和种群盛衰的趋势。     

中国乌蜂虻亚科二新种记述──（双翅目：蜂虹科）

本文记述乌蜂虹亚科Ｕｓｉｉｎａｅ２新种,分别隶属于Ｕｓｉａ（乌蜂虹属）和Ｐａｒａｇｅｒｏｎ（拟驼蜂虹属）,并对有关属的特征及分类情况作了概述。     

山羊关节炎─脑炎前病毒的PCR检测

山羊关节炎─脑炎前病毒的ＰＣＲ检测相文华,丁恩雨,秦运安,吕晓玲,沈荣显（中国农业科学院哈尔滨兽医研究所哈尔滨１５０００１）关键词聚合酶链反应,山羊关节炎—脑炎病毒,整合山羊关节炎—脑炎（ＣａｐｒｉｎｅＡｒｔｈｒｉｔｉｓ－Ｅｎｃｅｐｈａｌｉｔｉｓ,Ｃ...