TARGETED RANDOM MUTAGENESIS - THE USE OF AMBIGUOUSLY SYNTHESIZED OLIGONUCLEOTIDES TO MUTAGENIZE SEQUENCES IMMEDIATELY 5' OF AN ATG INITIATION CODON
TARGETED RANDOM MUTAGENESIS - THE USE OF AMBIGUOUSLY SYNTHESIZED OLIGONUCLEOTIDES TO MUTAGENIZE SEQUENCES IMMEDIATELY 5' OF AN ATG INITIATION CODON
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DOI：
10.1093/nar/11.10.3113
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发表时间：
1983-01-01
影响因子：
14.9
通讯作者：
HEYNEKER, HL
中科院分区：
文献类型：
--
作者：
MATTEUCCI, MD;HEYNEKER, HL
牛生长激素 (BGH) 结构基因起始密码子 5' 附近的 9 个碱基对已被诱变。诱变方法采用将模糊合成的寡核苷酸双链体连接到 BGH 表达质粒中先前设计的间隙中。突变方法与杂交筛选相结合，可有效分离目标区域内的 1 和 2 个碱基对变化。检测含有突变质粒的大肠杆菌培养物的 BGH 表达的相对水平。最显着的结果是在该区域的不同位置将 G 替换到 mRNA 中会产生不同的影响
The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) structural gene have been mutagenized. The mutagenesis method employs the ligation of an ambiguously synthesized oligonucleotide duplex into a previously engineered gap in an expression plasmid for BGH. The mutation method, coupled with hybridization screening, is efficient at isolating 1 and 2 base pair changes within the targeted region. The E. coli cultures harboring the mutant plasmids were assayed for relative leveLs of BGH expression. The most notable result is the varied effect of substitution of G into the mRNA at various positions in this region