人血红蛋白α、β基因以可溶形式在大肠杆菌中的表达

SOLUBLE EXPRESSION OF α AND β GENES OF HUMAN HEMOGLOBIN IN ESCHERICHIA COLI

将人血红蛋白α、β珠蛋白基因分别克隆进 pET 2 1b原核表达载体中 ,宿主菌经IPTG化学诱导 ,在1.6× 10 5处有特异的蛋白带表达。表达产物以可溶形式存在 ,α珠蛋白的表达量达总菌体蛋白的 5 % ,β珠蛋白的表达量达 15 % ,并经Western Blotting杂交证实。

Fragments of α and β genes were cloned into expression plasmid pET-21b. Bacteria harbouring expression plasmid was induced with IPTG,and a band of expressed protein of 16 kD was found in PAGE. They expressed as soluble products. α expression product reached about 5% of total bacterial protein,β expression product reached about 15% of total bacterial protein. The products were confirmed by Western-blotting.