Enhancement of heterologous polypeptide expression by alterations in the ribosome-binding-site sequence
Abstract In our studies to improve the expression of bovine growth hormone in E. coli , we have observed that the production of the hormone can be greatly increased by three similar AT-rich ribosome-binding-site sequences. Thus the low level expression of the hormone at less than 1% of the total cellular protein can be enhanced to about 15–30% by simply enriching the sequence flanking the Shine-Dalgarno site with adenine- and thymine-containing deoxyribonucleotides. These AT-rich ribosome-binding-site sequences enhance the expression of porcine growth hormone in a similar manner. These sequences also increase the moderate expression (about 5%) of tumour necrosis factor to high levels of about 30%.