人c-myc外显子1编码蛋白在大肠杆菌中的表达与表征

Miltiades C. Psallidopoulos , Arun Seth , Garrett C. Dubois , Robert J. Fisher , Takis S. Papas
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引用次数: 4

摘要

我们之前报道的c-myc基因座DNA序列研究数据表明,人类c-myc外显子1具有开放的重载框架,能够编码188个氨基酸残基的蛋白质。为了证实开放阅读框的存在,我们构建了一种重组载体(pMCP60),该载体包含λcII翻译起始区的一段、v-mos基因N末端的一部分和人类c-myc基因第一外显子的639个碱基对。pMCP60表达通过高压液相色谱纯化的38千道尔顿的三artate蛋白(cII-mos-myc)。通过部分氨基酸序列分析证实了cII-mos-myc融合蛋白中myc外显子1序列的存在。这些实验进一步证实,人类c-myc基因的第一个外显子包含一个能够在大肠杆菌中表达蛋白质的开放阅读框。
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Expression and characterization of a protein encoded by the human c-myc exon 1 in Eschirichia coli

Our previously reported data from DNA sequence studies of the c-myc locus show that the human c-myc exon 1 has an open reaading frame capable of encoding a protein of 188 amino acid residues. To confirm the presence of the open reading frame, we constructed a recombinant vector (pMCP60) that contains a segment of the λ cII translational initiation region, a portion of the N-terminus of the v-mos gene, and 639 base pairs of the first exon of the human c-myc gene. pMCP60 expresses a 38 kilodalton tripartate protein (cII-mos-myc), which was purifed by high-pressure liquid chromatography. The presence of myc exon 1 sequences in the cII-mos-myc fusion protein was confirmed by partial amino acid sequence analysis. These experiments further establish that the first exon of the human c-myc gene contains an open reading frame capable of expressing a protein in Escherichia coli.

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