杂交基因和蛋白质的生产方法及其在评估同源色氨酸合成酶α多肽中氨基酸差异的重要性方面的应用。
Procedure for production of hybrid genes and proteins and its use in assessing significance of amino acid differences in homologous tryptophan synthetase alpha polypeptides.
作者信息
Schneider W P, Nichols B P, Yanofsky C
出版信息
Proc Natl Acad Sci U S A. 1981 Apr;78(4):2169-73. doi: 10.1073/pnas.78.4.2169.
Abstract
Hybrid tryptophan synthetase alpha and beta polypeptides were produced by genetic recombination between the trpB--trpA regions of Escherichia coli and Salmonella typhimurium contained on compatible, multicopy plasmids. Intragenic recombination was decreased but still evident in recA cells. Genetic exchange occurred at many sites within trpA, but every recombinant gene produced a functional alpha polypeptide despite many amino acid differences from one or the other of the parental polypeptides. The five hybrid tryptophan synthetase alpha subunits examined resembled the parental polypeptides in catalytic function but differed in thermostability. The stability differences suggest that, as amino acid changes occurred in these proteins during the course of evolution, subsequent changes were limited to those that would allow retention of a desired protein conformation.
摘要
通过位于相容多拷贝质粒上的大肠杆菌和鼠伤寒沙门氏菌的trpB-trpA区域之间的基因重组，产生了杂合色氨酸合成酶α和β多肽。在recA细胞中，基因内重组减少但仍然明显。基因交换发生在trpA内的许多位点，但每个重组基因都产生了一种功能性α多肽，尽管与亲本多肽中的一种或另一种有许多氨基酸差异。所检测的五个杂合色氨酸合成酶α亚基在催化功能上类似于亲本多肽，但在热稳定性上有所不同。稳定性差异表明，在进化过程中这些蛋白质发生氨基酸变化时，随后的变化仅限于那些能够保持所需蛋白质构象的变化。
相似文献
1
Procedure for production of hybrid genes and proteins and its use in assessing significance of amino acid differences in homologous tryptophan synthetase alpha polypeptides.杂交基因和蛋白质的生产方法及其在评估同源色氨酸合成酶α多肽中氨基酸差异的重要性方面的应用。
Proc Natl Acad Sci U S A. 1981 Apr;78(4):2169-73. doi: 10.1073/pnas.78.4.2169.
2
3
Fusion of trpB and trpA of Escherichia coli yields a partially active tryptophan synthetase polypeptide.大肠杆菌的trpB和trpA融合产生一种部分活性的色氨酸合成酶多肽。
J Biol Chem. 1987 Aug 25;262(24):11584-90.
4
Structure and properties of a hybrid tryptophan synthetase of alpha chain produced by genetic exchange between Escherichia coli and Salmonella typhimurium.通过大肠杆菌和鼠伤寒沙门氏菌之间的基因交换产生的α链杂合色氨酸合成酶的结构与特性
Proc Natl Acad Sci U S A. 1977 Jan;74(1):286-90. doi: 10.1073/pnas.74.1.286.
5
Cloning and sequence analysis of tryptophan synthetase genes of an extreme thermophile, Thermus thermophilus HB27: plasmid transfer from replica-plated Escherichia coli recombinant colonies to competent T. thermophilus cells.嗜热栖热菌HB27色氨酸合成酶基因的克隆与序列分析：从影印平板培养的大肠杆菌重组菌落向感受态嗜热栖热菌细胞的质粒转移
J Bacteriol. 1990 Jun;172(6):3490-5. doi: 10.1128/jb.172.6.3490-3495.1990.
6
On the role of helix 0 of the tryptophan synthetase alpha chain of Escherichia coli.关于大肠杆菌色氨酸合成酶α链螺旋0的作用。
J Biol Chem. 1996 Jun 21;271(25):14754-63. doi: 10.1074/jbc.271.25.14754.
7
Relative activities and stabilities of mutant Escherichia coli tryptophan synthase alpha subunits.突变型大肠杆菌色氨酸合成酶α亚基的相对活性和稳定性
J Bacteriol. 1991 Mar;173(6):1886-93. doi: 10.1128/jb.173.6.1886-1893.1991.
8
Directed evolution of (βα)(8)-barrel enzymes: establishing phosphoribosylanthranilate isomerisation activity on the scaffold of the tryptophan synthase α-subunit.（βα）（8）桶状酶的定向进化：在色氨酸合酶α亚基的支架上建立磷酸核糖基氨基苯甲酸异构酶活性。
Protein Eng Des Sel. 2012 Jun;25(6):285-93. doi: 10.1093/protein/gzs015. Epub 2012 Apr 6.
9
PCR mutagenesis and overexpression of tryptophan synthase from Salmonella typhimurium: on the roles of beta2 subunit Lys-382.鼠伤寒沙门氏菌色氨酸合酶的PCR诱变与过表达：关于β2亚基赖氨酸-382的作用
Protein Expr Purif. 1996 Aug;8(1):126-36. doi: 10.1006/prep.1996.0082.
10
Site-directed mutagenesis of the alpha subunit of tryptophan synthase from Salmonella typhimurium.
Biochem Biophys Res Commun. 1988 Mar 15;151(2):672-8. doi: 10.1016/s0006-291x(88)80333-4.
引用本文的文献
1
Establishing wild-type levels of catalytic activity on natural and artificial (beta alpha)8-barrel protein scaffolds.在天然和人工（β-α）8桶蛋白支架上建立野生型催化活性水平。
Proc Natl Acad Sci U S A. 2009 Mar 10;106(10):3704-9. doi: 10.1073/pnas.0810342106. Epub 2009 Feb 23.
2
Directed evolution of a (beta alpha)8-barrel enzyme to catalyze related reactions in two different metabolic pathways.对一种（β-α）8桶状酶进行定向进化，以催化两条不同代谢途径中的相关反应。
Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):9925-30. doi: 10.1073/pnas.160255397.
3
Mutational analysis of the active site of indoleglycerol phosphate synthase from Escherichia coli.大肠杆菌磷酸吲哚甘油合酶活性位点的突变分析。
Protein Sci. 1998 May;7(5):1221-32. doi: 10.1002/pro.5560070518.
4
Gene conversion tracts from double-strand break repair in mammalian cells.哺乳动物细胞双链断裂修复产生的基因转换序列。
Mol Cell Biol. 1998 Jan;18(1):93-101. doi: 10.1128/MCB.18.1.93.
5
Structural similarity between ornithine and aspartate transcarbamoylases of Escherichia coli: implications for domain switching.大肠杆菌鸟氨酸转氨甲酰酶和天冬氨酸转氨甲酰酶之间的结构相似性：对结构域切换的影响
Protein Sci. 1996 Apr;5(4):719-28. doi: 10.1002/pro.5560050417.
6
Molecular evolution of the Escherichia coli chromosome. IV. Sequence comparisons.大肠杆菌染色体的分子进化。IV. 序列比较。
Genetics. 1993 Mar;133(3):455-68. doi: 10.1093/genetics/133.3.455.
7
Genetic map of Salmonella typhimurium, edition VIII.鼠伤寒沙门氏菌遗传图谱，第八版。
Microbiol Rev. 1995 Jun;59(2):241-303. doi: 10.1128/mr.59.2.241-303.1995.
8
Trp aporepressor production is controlled by autogenous regulation and inefficient translation.色氨酸阻遏物的产生受自身调节和低效翻译的控制。
Proc Natl Acad Sci U S A. 1982 May;79(10):3120-4. doi: 10.1073/pnas.79.10.3120.
9
The complete nucleotide sequence of the tryptophan operon of Escherichia coli.大肠杆菌色氨酸操纵子的完整核苷酸序列。
Nucleic Acids Res. 1981 Dec 21;9(24):6647-68. doi: 10.1093/nar/9.24.6647.
10
A ribosome binding site sequence is necessary for efficient expression of the distal gene of a translationally-coupled gene pair.核糖体结合位点序列对于翻译偶联基因对中远端基因的高效表达是必需的。
Nucleic Acids Res. 1984 Jun 11;12(11):4757-68. doi: 10.1093/nar/12.11.4757.
本文引用的文献
1
Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium.大肠杆菌和鼠伤寒沙门氏菌中色氨酸合成酶B基因的核苷酸序列。
J Mol Biol. 1980 Oct 5;142(4):489-502. doi: 10.1016/0022-2836(80)90259-4.
2
Sequencing end-labeled DNA with base-specific chemical cleavages.通过碱基特异性化学切割对末端标记的DNA进行测序。
Methods Enzymol. 1980;65(1):499-560. doi: 10.1016/s0076-6879(80)65059-9.
3
Comparison of the tryptophan synthetase alpha-subunits of several species of Enterobacteriaceae.几种肠杆菌科细菌色氨酸合成酶α亚基的比较。
J Bacteriol. 1966 May;91(5):1819-26. doi: 10.1128/jb.91.5.1819-1826.1966.
4
Immunochemical and enzymatic comparisons of the tryptophan synthase alpha subunits from five species of Enterobacteriaceae.五种肠杆菌科细菌色氨酸合成酶α亚基的免疫化学和酶学比较
J Bacteriol. 1969 Mar;97(3):1310-20. doi: 10.1128/jb.97.3.1310-1320.1969.
5
The functional significance of the evolutionary divergence between the tryptophan operons of Escherichia coli and Salmonella typhimurium.大肠杆菌和鼠伤寒沙门氏菌色氨酸操纵子之间进化差异的功能意义。
J Mol Evol. 1974 Nov 29;4(2):121-37. doi: 10.1007/BF01732018.
6
Construction of biologically functional bacterial plasmids in vitro.体外构建具有生物学功能的细菌质粒。
Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240-4. doi: 10.1073/pnas.70.11.3240.
7
The use of thin acrylamide gels for DNA sequencing.用于DNA测序的薄丙烯酰胺凝胶的使用。
FEBS Lett. 1978 Mar 1;87(1):107-10. doi: 10.1016/0014-5793(78)80145-8.
8
Nucleotide sequences of trpA of Salmonella typhimurium and Escherichia coli: an evolutionary comparison.鼠伤寒沙门氏菌和大肠杆菌色氨酸合成酶A亚基的核苷酸序列：进化比较
Proc Natl Acad Sci U S A. 1979 Oct;76(10):5244-8. doi: 10.1073/pnas.76.10.5244.
9
Extracellular nucleases of pseudomonas BAL 31. III. Use of the double-strand deoxyriboexonuclease activity as the basis of a convenient method for the mapping of fragments of DNA produced by cleavage with restriction enzymes.铜绿假单胞菌BAL 31的细胞外核酸酶。III. 以双链脱氧核糖核酸外切酶活性为基础，建立一种便捷的方法用于绘制由限制性内切酶切割产生的DNA片段图谱。
Nucleic Acids Res. 1978 May;5(5):1445-64. doi: 10.1093/nar/5.5.1445.
10
Structural and functional analysis of cloned deoxyribonucleic acid containing the trpR-thr region of the Escherichia coli chromosome.含大肠杆菌染色体trpR-thr区域的克隆脱氧核糖核酸的结构与功能分析。
J Bacteriol. 1979 Oct;140(1):106-13. doi: 10.1128/jb.140.1.106-113.1979.
Zotero 插件