反义技术研究进展

过去几年目睹了反义技术的爆炸性发展,文章较全面地评述了反义技术的主要内容和近几年来的研究进展,它们包括几类受人注意的DNA类似物、RNA和Ribozyme.。此外,基于研究现状还展示了反义技术的前景。     

利用~1H-NMR模拟谱研究二核苷酸A3′P(CH_3)5′A的构象性质

本文根据Altona等人的方法,在利用~1H-NMR模拟谱确定18℃、40℃、70℃三种温度下有关质子化学位移及偶合常数的基础上,分析了A3′P(CH_3)5′A的两种非对映异构体a和b的构象状态。它们与A3′P(OH)5′A相比发现:(1)在18℃时两个核糖环是S型构象占主要成分,并随温度升高S型构象成分增多;(2)磷酸骨架的扭角ε′和β′分别改变±15°及±4°;(3)在优势构象情况下,两个核糖环都部分重叠,而且A3′P(CH_3)5′A(a)的重叠程度比A3P′(CH_3)5′A(b)小;(4)腺嘌呤碱基都更倾向去堆积状态,而且随温度升高A3′P(CH_3)5′A(a)比A3′P(CH_3)5′A(b)有着更强的去堆积效应。     

用anti-RNA遏制要不得的基因

anti-RNA——研制者称之为反(anti-)mRNA、反义(anti-sense)RNA、     

DNA启动子的计算机识别

本文介绍了两种预测DNA顺序上的启动子位置的计算机识别方法,方法1是基于启动子部位单核苷酸的分布不均一性,方法2是基于启动子部位二核苷酸的分布不均一性。分别用这两种方法推测了质粒pBR322DNA上启动子的位置,从而验证了化学实验的结果。     

用改进的寡核苷酸诱导突变法改造人胰岛素前体基因

我们用改进了的寡聚核苷酸诱导突变法,将两个单一限制性内切酶位点引入人胰岛素前体B链与C链连接处附近,及C链与A铁连接处附近。用含U模板法将B链第30个残基密码子ACC改成ACG,从而引入MluⅠ位点。用修改了的缺口双链DNA突变法,将A链第4个残基密码于GAG改成CTG,引入了一个PstⅠ位点。突变效率的为17％-36％。这个突变体在人胰岛素前体结构及蛋白质折叠的研究中,将有利于更换不同的C-肽。     

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)