PRODUCTION OF UNMODIFIED HUMAN ADULT HEMOGLOBIN IN ESCHERICHIA-COLI
PRODUCTION OF UNMODIFIED HUMAN ADULT HEMOGLOBIN IN ESCHERICHIA-COLI
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DOI：
10.1073/pnas.90.17.8108
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发表时间：
1993-09-01
影响因子：
11.1
通讯作者：
HO, C
中科院分区：
文献类型：
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作者：
SHEN, TJ;HO, NT;HO, C
我们已经构建了一个质粒（pHE 2），其中合成的人α-和β-珠蛋白基因和甲硫氨酸氨基肽酶（Met-AP）基因从大肠杆菌的控制下，单独的tac启动子共表达。在E. coliJM 109，经快速蛋白液相色谱纯化，得到两个主要组分a和B。电喷雾质谱法显示，至少98%和约90%的组分a的表达的α和β链分别具有预期质量。组分a中剩余10%的β链在质量上对应于β链加甲硫氨酸。在组分B中，α和β链都具有正确的质量，没有可检测的N-末端甲硫氨酸（
We have constructed a plasmid (pHE2) in which the synthetic human alpha- and beta-globin genes and the methionine aminopeptidase (Met-AP) gene from Escherichia coli are coexpressed under the control of separate tac promoters. The Hbs were expressed in E. coli JM109 and purified by fast protein liquid chromatography, producing two major components, a and b. Electrospray mass spectrometry shows that at least 98% and about 90% of the expressed alpha and beta chains of component a, respectively, have the expected masses. The remaining 10% of the beta chain in component a corresponds in mass to the beta chain plus methionine. In component b, both alpha and beta chains have the correct masses without detectable N-terminal methionine (