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MODIFIED PHEUMOLYSINE RECOMBINANT PLASMID, METHOD OF PRODUCING MODIFIED PHEUMOLYSINE AND VACCINE

机译：修饰的磷脂酰赖氨酸重组质粒，修饰的磷脂酰赖氨酸和疫苗的生产方法

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FIELD: gene engineering, more particularly preparation of modified pneumolysine and manufacture of vaccine based thereon. SUBSTANCE: modified pneumolysine is produced by oligonucleotide directed mutagenesis. Modified wild-type pneumolysine gene is then expressed in bacteria E.Coli. Modified pneumolysine has certain amino acid sequence. One or more amino acid replacements, deletions or blocking mutations are made in sites 427-437, 367-398 and 257-297 of said amino acid sequence. Recombinant plasmid that determines synthesis of modified pneumolysine is constructed by vector pKK233-2 and comprises ampicillin- and tetracycline-resistant genes. Isopropyl -D- thiogalactopyranoside regulated promoter trc. Ribosom lac binding site adjacent to initiation codon ATG. Restriction sites for endonulceases Pst I and Hind III. Modified pneumolysine gene cloned into NCO I site which has certain sequence of nucleotides. Vaccine capable of inducing immune response in animals comprises antigen from modified pneumolysine. EFFECT: improved properties of the vaccine. 7 cl
机译：领域：基因工程，更具体地讲，修饰的肺炎赖氨酸的制备和基于其的疫苗的制备。物质：修饰的肺炎赖氨酸是通过寡核苷酸定向诱变产生的。然后在细菌大肠杆菌中表达修饰的野生型肺炎球菌赖氨酸基因。修饰的肺炎赖氨酸具有一定的氨基酸序列。在所述氨基酸序列的位点427-437、367-398和257-297中进行一种或多种氨基酸置换，缺失或阻断突变。通过载体pKK233-2构建可确定修饰的肺炎赖氨酸合成的重组质粒，该质粒包含氨苄青霉素和四环素抗性基因。异丙基-D-硫代吡喃半乳糖苷调节的启动子trc。与起始密码子ATG相邻的核糖核酸lac结合位点。内切酶Pst I和Hind III的限制性酶切位点。修饰的肺炎球菌赖氨酸基因克隆到具有一定核苷酸序列的NCO I位点。能够在动物中诱导免疫应答的疫苗包含来自修饰的肺炎赖氨酸的抗原。效果：改善了疫苗的性能。 7厘升

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