用ST-EPR研究胆酸对F_1-F_0在F_1-F_0复合体中慢运动的影响

本文比较了经胆酸处理和对照样品的马未酰亚胺自旋标记的F_1-F_0酶复合体的ST-EPR波谱, 二者波谱呈现明显不同的形态.经胆酸处理的复台体中F_h-F_0酶蛋白的波谱参数L”/L’,C’/C,H”/H较对照样品明显增大.选用参数L”/L’计算其相关时间τ_c,得出对照的F_1-F_0酶蛋白的τ_e为5.5×10~(-5)秒而经胆酸处理的F_1-F_0酶蛋白的τ_e为1×10~(-4)秒.表明胆酸处理的样品中酶蛋白运动大大减慢.因此推测胆酸可能通过增加F_1-F_0酶蛋白在膜脂中的旋转运动相关时间,减慢其运动速率,来抑制Mg~2 对F_1-F_0酶水解活力的激活作用的.     

应用酶联免疫吸附试验检测不同类型乙型肝炎中激活补体类HBsAg循环免疫复合物

本文以抗人C_(?)的羊IgG为包被抗体,以HRP-HBs抗体为指示抗体,建立了可检测激活补体类HBsAg循环免疫复合物(HBsAg/C3-CIC)的C_3捕捉法酶联免疫吸附试验。检测了236例六种类型临床诊断为乙型肝炎的病人血清标本,其阳性率分别为:无症状携带者(ASC)12.9％(4/31),急性肝炎(AH)36.7％(22/60),慢性迁延性肝炎(CPH)33.3％(7/21),慢性活动性肝炎(CAH)59.6％(34/57),重型肝炎(SH)77.8％(14/18),肝炎后肝硬化(PLC)67.3％(33/49),阳性率与肝损严重程度明显相关(P<0.01)。认为HBs-Ag/C3-CIC可能在乙型肝炎病毒引起的慢性活动性肝炎、重型肝炎和肝炎后肝硬化的发病过程中起重要作用,并可作为乙型肝炎的诊断、临床分型和预后判断的指标之一。     

恒河猴B病毒相关抗体检测方法的比较

以NT方法为基础比较了ELISA和FIA方法,共检测84份猴B病毒相关抗体的敏感性,结果ELISA和EIA阳性各50份(59.5％),NT阳性45份(53.6％)。三种方法相符者71份,符合率84.5％。ELISA和FIA均较NT敏感,而且快速,简便,经济,可应用于大批标本的检查。     

血液病患者的巨细胞病毒感染

用间接酶联免疫吸附实验(ELISA)对新近诊断的179例血液病患者血清巨细胞病毒(HCMV)IgM和IgA抗体进行了检测。阳性率分别为11,17％11,73％,明显高于对照人群(4,76％和3,97％),提示血液病患者由于免疫功能下降,易于发生HCMV活动性感染。     

HSV在两种细胞培养上抗原出现规律(的观察)及细胞敏感性的比较

本实验观察比较了Hep—2细胞和兔肾细胞对单纯疱疹病毒的敏感性。用免疫荧光法和免疫酶标法检测了细胞培养上HSV抗原出现的规律。实验结果指出:Hep—2细胞和兔肾细胞对HSV的敏感性一致;接种病毒后6小时,可以在Hep—2细胞和兔肾细胞上发现HSV抗原;免疫荧光法和免疫酶标法是进行病毒学研究和病毒感染早期诊断的可靠方法。     

大肠杆菌棉子糖操纵子a-半乳糖苷酶表达的调节控制

大肠杆菌棉子糖操纵子(raf)位于质粒,其第一结构基因rafA编码的a-半乳糖苷酶为诱导酶。Rat操纵子比乳糖操纵子(lac)或蜜二糖操纵子(mel)对诱导物有更严格的结构特异性。该酶被蜜二糖或棉子糖诱导,也被D-半乳糖微弱诱导,但不受乳糖、PNPG等结构相近糖所诱导。A-半乳糖苷酶的酶诱导形成能力在对数生长末期出现高峰。Rat 操纵子基因结构组成及调节与乳糖操纵子相似。以阻遏物为中介的负调控在raf操纵子调节中起主要作用,同时以环腺苷一代谢降解物基因激活蛋白(cAMP-CAP)为中介的正调控也参与调节。当0．4％葡萄糖加入到其它碳源培养基时,该酶表达水平下降至原活力的1／2—1／3。无论诱导或组成型酶的葡萄糖抑制均未见瞬时抑制。腺苷环化酶(cya)缺失或环腺苷受体蛋白(crP)和cya双缺陷菌株的酶表达则分别下降到原活力的9％和2．5％。Cya突变株或葡萄糖对raf操纵子表达的抑制可被cAMP解除,但cya和crP双缺陷菌株仍有葡萄糖抑制,而且这种抑制不为cAMP抵消,表明通过降低cAMp而影响cAMP-CAP复合体形成还不能解释代谢降解物抑制的全部机制。尚无证据说明吲哚类小分子化合物和低浓度尿素对raf操纵子表达的明显作用。     

大肠杆菌棉子糖操纵子α—半乳糖苷酶表达的调节控制

The alpha-galactosidase, coded for by the first structural gene rafA in the plasmid determined raf operon was an inducible enzyme. In contrast to lac or mel operon, raf operon has more strict structural specificity for inducers. The enzyme can be induced by melibiose and raffinose, or weakly by D-galactose, but not by structurally related sugars such as lactose, PNPG etc.. The alpha-galactosidase forming capacity as function of growth curve reached a single peak at the end of the logarithmic phase of the growth. The structure and regulation of raf operon is similar to those of lac operon. The repressormor-mediated negative control plays a major role in the regulation of raf operon, and cAMP-CAP mediated positive control is also involved in the regulation. When 0.4% glucose was added into the medium with other carbon sources, the expression of the enzyme was repressed by 2-3 fold. Transient catabolite repression has been observed neither in inducible nor constitutive alpha-galactosidase expression. Based on alpha-galactosidase assay, in mutant strains CA8306(cya) and CA8445 (cya, crp) the expression level of raf operon was only 9% and 2.5% of that in wild type strain respectively. The glucose effect or the repression in cya mutant can be abolished by 1-5 mmol cAMP. The constitutive alpha-galactosidase expression in cya and cry double mutant (CA8445) remains repressible by glucose, but irreversible by cAMP, suggesting cAMP-CAP complex is not the exclusive mediator of the catablite repression.(ABSTRACT TRUNCATED AT 250 WORDS)     

大肠杆菌青霉素G酰化酶基因及其邻近区域的核苷酸全序列

Some of microorganisms have been known to possess penicillin G acylase activity. The E. coli derived penicillin G acylase (PGA) can catalyze the conversion of penicillin G into phenylacetic acid and 6-amino-penicillanic acid, the latter is used as the starting compound for the industrial formation of semi-synthetic penicillins. Apart from its industrial importance, the enzyme PGA displays a number of interesting properties. Catalytically active enzyme is localized in the periplasmic space of E. coli cells and composed of two dissimilar subunits. The two subunits are apparently produced from a precursor protein, via a processing pathway hitherto unique in its features for a prokaryotic enzyme. The studies on processing of the precursor and on the relationship between structure and function of the mature enzyme are important theoretically. Previously we cloned a 3.5 kb DNA fragment from a strain (E. coli AS 1.76), which displays PGA activity. In this paper, we report a nucleotide sequence of the 3.5 kb DNA fragment containing PGA gene. After insertion of the DNA fragment into EcoR I and Hind III sites in pWR 13, pPGA 20 had been obtained. We subcloned the Hind III and Bg1 II treated fragment of 1.6 kb in length from pPGA 20 into Hind III and BamH I sites of pWR 13 to get a pPGA 1.6, and Bg1 II and EcoR I treated fragment of 1.9 kb in length into BamH I and EcoR I sites of pWR 13 to get a pPGA 1.9. The linearized pPGA 1.9 which were digested with appropriate restriction enzymes were progressively shortened from both ends respectively by digestion with Bal 31 nuclease, followed by cleavage of shortened target DNA off vector DNA molecules with appropriate restriction enzymes. The series of the DNA fragments shortened from EcoR I end were then cloned into plasmid pWR 13 which had previously digested with Hind III and Sma I enzymes (Fig. 1). The DNA fragment cloned in pWR 13 were directly sequenced on the resulted plasmids by using primer I and primer II. Thus we have obtained the complete nucleotide sequence of the 3.5 kb DNA fragment. The 3.5 kb fragment contains an intact PGA gene which is 2.6 kb.(ABSTRACT TRUNCATED AT 400 WORDS)