中国生物工程杂志

Adiponectin, as a specific adipokine, entails diverse biological roles associated with obesity, insulin resistance, typeⅡdiabetes and cardiovascular diseases, with broad potentials of application. In this study, by using the total human blood genomic DNA as template, we particularly exploited the approach of splicing by overlap extension (SOE) to combine the two individually amplified exon fragments containing adiponectin-coding regions, and obtained the accurate, full-length of adiponectin gene (AP) verified by sequencing analysis. Subsequently, an expression construct pET-wAP was achieved by fusing the adiponectin gene behind the thioredoxin region within the potent plasmid pET32a(+). The fusion form of thioredoxin-adiponectin was successfully expressed by IPTG induction, with a high yield up to 41% of the total cell protein but mainly in inclusion bodies. Further studies on efficient and soluble expression of human adiponectin and its globular domain are now in proceeding.