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)     

光合作用反应中心研究的一个重大成就：1988年度诺贝尔化学奖获奖项目简介

由于光合作用反应中心三度空间结构的阐明,使三位在西德工作的科学家获得了1988年度诺贝尔化学奖。他们是约翰·戴森霍弗尔(Johann·Deisenhofer);罗伯特·休贝尔(Robert·Huber)和哈特穆特·米歇尔(Hartmut·Michel)。这是继卡尔文(M.Calvin)研究光合二氧化碳同化途径获得1961年度诺贝尔化学奖以来,另一个直接授予光合作用研究成就的诺贝尔奖。瑞典皇家科学院在颁布授奖公报中指出:“他们第一次成功地阐明膜蛋白形成的详细过程,并从原子水平上揭示了光合作用反应中心由原子形成的分子结构”。这一重大成就标志着光     

甘肃盐池湾白唇鹿自然保护区有蹄类动物群落结构的初步观察

1987年8—10月,作者在甘肃省肃北蒙古族自治县盐池湾白唇鹿自然保护区研究了有蹄类动物栖息地类型、群落结构和数量等级。     

酵母ENO2上游激活顺序对酵母 LEU2表达的影响

本文将酿酒酵母(Saccharomyces cerevisiae)中编码烯醇化酶的基因之—ENO2的上游墩活顺序嵌入穿梭质粒YEpl3上酵母LEU2基因上游—405Hpa I的酶切部位。LEU2为编码β-异丙基苹果酸脱氢酶基因。从而研究了酵母ENO2的上游激活顺序对酵母LEU2表达的影响。实验结果表明ENO2上游激活顺序不论正向或反向嵌八都激活LEU2的表达达四倍左右。有亮氨酸存在的条件下,LEU2的表达受到抑制。ENO2上游激活顺序在对LEU2表达的激活上并不受葡萄糖的诱导。提出了用ENO2上游激活顺序组建高表达系统的可能性。     

小麦高分子量谷蛋白亚基基因片段的体外扩增

生物体尤其是高等动植物,绝大多数基因的拷贝数是很低的,如果没有基因扩增的手段,几乎无法研究基因的结构及其与表达的关系。当今分子生物学发展中最基本的技术之一基因克隆,实际上就是基因体内扩增(in vivo amplification)的技术。近年来发展了一种基因体外扩增(in vitro amplification)的新技术,又称     

几种固氮菌nifA基因片段的同源性分析

参照已知数种固氮菌nifA基因的DNA序列,选择其中间区域的两个保守性较强的序列合成引物,利用肺炎克匠杆菌(Klebsiella pneumoniaef)、棕色固氮菌(Azotobacter vinela-ndii)、巴西固氮螺菌(Azospirllum brasilense)、草螺菌(Herbaspirillum seropedicae)和深红红螺菌(Rhodospirillum rubrum)的总DNA进行聚合酶链反应,结果均扩增出约450bp大小的片段,经证实为各种固氮菌的nifA基因部分片段。 核酸印迹分子杂交结果显示出nifA基因在不同固氮菌中的同源性不强。     

海南大田自然保护区海南坡鹿种群动态研究

笔者自1986年起对仅存于海南大田国家级保护区围栏内外两个坡鹿种群进行了连续4年追踪研究,就数量、年净增长率、年均增长率、年龄组成、性比、成幼比、母仔比、及补充率和死亡率进行了分析。围栏内外坡鹿年均增长率22.5％和10.6％,年净增长率逐年呈下降趋势。补充率下降是围栏内坡鹿年净增长率下降的主要原因,而这是由于密度过高、食物不足造成的;围栏外坡鹿年净增长率下降是死亡率增加的结果,偷猎是主要因素。对性比、成幼比、母仔比变化的分析也得出了同样的结论。本文最后提出了保护建议。