Alikhanian S I
Genetika. 1976;12(7):150-73.
The review of literature (1970-1976) on problems of gene engineering is given. Gene engineering is pointed out to be a new method of modern biology and a new page of modern molecular genetics. Gene engineering detected a real possibility of artificial creating living hybrid organisms, i.e. constructing functional recombinant DNA molecules according to a project of investigator, but not to possibilities of crossing. The determination of gene engineering (in contrast with genetical engineering) is given in the first division of the article. Genetical engineering is a construction of hybrid organisms on the basis of recombination between non-homologous chromosomes cy crossing. Genetical engineering is based on sex crossing, thus the application of this method is restricted by crossability (i.e. experiments in vivo), which possibilities are determined by taxonomical limits. Gene engineering is a new method of operating directly with genes. It permits constructing in vitro any hybrid genomes desirable. There is no limits of combining ability for gene engineering. Three main stages of constructing hybrid genomes should be taken into account for the proper determination of gene engineering as a method of genome constructing: 1) the gene isolation; 2) their cross-linking in vitro; 3) the transfer of hybrid DNA into recipient cell or its genome. The cardinal stage of gene engineering is the construction of hybrid DNA, cross-linking any initial DNAs from any remote animals, plants and bacteria. All the methods known of gene isolation are described. The chemical method of gene isolation is based on that case, when DNA of some gene differs in its physico-chemical characteristics from total DNA, for example, DNAs of genes coding ribosomal RNAs or sea urchine histone DNA. Isolation of promotors and operators using DNA dependent RNA polymerase, which recognizes promotors, repressor and operator DNA, should also be considered as the chemical method of gene isolation. Restrictase method, which is also well known, is convenuent because the restricts have long enough sticky ends, which is important for the following gene cross-linking. The method of total restriction, reported by Lederberg et al. and Debabov et al., is described. The phage method (in particular, Shimada method) is given, permitting the direct integration of lambda phage into a number of sites of Escherichia coli chromosome. Gene engineering method of gene isolation is mentioned, in particular, the data of Kameron et al. on hybrid phages carrying DNA ligase gene, and Clark a. Carbon on hybrid plasmids carrying triptophane and arabinose operons genes. These methods are called "shot gun". Methods of gene isolation from higher organisms are less developed. A method of gene isolation using so called colony hybridization (according to Grünstein and Hognes) is also given...
本文对1970 - 1976年有关基因工程问题的文献进行了综述。基因工程被指出是现代生物学的一种新方法,也是现代分子遗传学的新篇章。基因工程揭示了人工创造活的杂交生物的真正可能性,即根据研究者的设计构建功能性重组DNA分子,而非通过杂交的方式。文章第一部分给出了基因工程(与遗传工程相对)的定义。遗传工程是基于非同源染色体间的重组通过杂交构建杂交生物。遗传工程基于有性杂交,因此该方法的应用受杂交亲和性(即在体内进行实验)的限制,其可能性由分类学界限决定。基因工程是一种直接对基因进行操作的新方法。它允许在体外构建任何所需的杂交基因组。基因工程不存在组合能力的限制。为了恰当地将基因工程定义为一种基因组构建方法,应考虑构建杂交基因组的三个主要阶段:1)基因分离;2)它们在体外的交联;3)将杂交DNA转移到受体细胞或其基因组中。基因工程的关键阶段是构建杂交DNA,即将来自任何远缘动物、植物和细菌的任何初始DNA进行交联。文中描述了所有已知的基因分离方法。基因分离的化学方法基于这样一种情况,即某些基因的DNA在物理化学特性上与总DNA不同,例如,编码核糖体RNA的基因的DNA或海胆组蛋白DNA。利用依赖DNA的RNA聚合酶识别启动子、阻遏物和操纵基因DNA来分离启动子和操纵基因,也应被视为基因分离的化学方法。众所周知的限制性内切酶方法很方便,因为限制性内切酶具有足够长的粘性末端,这对后续的基因交联很重要。文中描述了莱德伯格等人和德巴博夫等人报道的完全限制法。给出了噬菌体法(特别是岛田法),该方法允许λ噬菌体直接整合到大肠杆菌染色体的多个位点。文中提到了基因分离的基因工程方法,特别是卡梅伦等人关于携带DNA连接酶基因的杂交噬菌体的数据,以及克拉克和卡尔邦关于携带色氨酸和阿拉伯糖操纵子基因的杂交质粒的数据。这些方法被称为“鸟枪法”。从高等生物中分离基因的方法发展较少。文中还给出了一种利用所谓的菌落杂交(根据格伦斯坦和霍格尼斯)来分离基因的方法……