Single-step affinity and cost-effective purification of recombinant proteins using the Sepharose-binding lectin-tag from the mushroom Laetiporus sulphureus as fusion partner.

Previous research showed that a lectin from the mushroom Laetiporus sulphureus, designed LSL, bound to Sepharose and could be eluted by lactose. In this study, by taking advantage of the strong affinity of LSL-tag for Sepharose, we developed a single-step purification method for LSL-tagged fusion proteins. We utilized unmodified Sepharose-4B as a specific adsorbent and 0.2 M lactose solution as an elution buffer. Fusion proteins of LSL-tag and porcine circovirus capsid protein, designated LSL-Cap was recovered with purity of 90 ± 4%, and yield of 87 ± 3% from crude extract of recombinant Escherichia coli. To enable the remove of LSL-tag, tobacco etch virus (TEV) protease recognition sequence was placed downstream of LSL-tag in the expression vector, and LSL-tagged TEV protease, designated LSL-TEV, was also expressed in E. coli., and was recovered with purity of 82 ± 5%, and yield of 85 ± 2% from crude extract of recombinant E. coli. After digestion of LSL-tagged recombinant proteins with LSL-TEV, the LSL tag and LSL-TEV can be easily removed by passing the digested products through the Sepharose column. It is of worthy noting that the Sepharose can be reused after washing with PBS. The LSL affinity purification method enables rapid and inexpensive purification of LSL-tagged fusion proteins and scale-up production of native proteins.

Analysis of the seismic performance of isolated buildings according to life-cycle cost.

This paper proposes an indicator of seismic performance based on life-cycle cost of a building. It is expressed as a ratio of lifetime damage loss to life-cycle cost and determines the seismic performance of isolated buildings. Major factors are considered, including uncertainty in hazard demand and structural capacity, initial costs, and expected loss during earthquakes. Thus, a high indicator value indicates poor building seismic performance. Moreover, random vibration analysis is conducted to measure structural reliability and evaluate the expected loss and life-cycle cost of isolated buildings. The expected loss of an actual, seven-story isolated hospital building is only 37% of that of a fixed-base building. Furthermore, the indicator of the structural seismic performance of the isolated building is much lower in value than that of the structural seismic performance of the fixed-base building. Therefore, isolated buildings are safer and less risky than fixed-base buildings. The indicator based on life-cycle cost assists owners and engineers in making investment decisions in consideration of structural design, construction, and expected loss. It also helps optimize the balance between building reliability and building investment.

Assessment of organochlorine pesticide contamination in relation to soil properties in the Pearl River Delta, China.

High levels of organochlorine pesticides (OCPs) such as hexachlorocyclohexanes (HCHs) and dichlorodiphenyltrichloroethanes (DDTs) have been found in soil of the Pearl River Delta (PRD), attributable to high pesticide application in this area. Consequently, the occurrence and environmental effect of HCHs and DDTs in the PRD have attracted considerable attention. However, study focusing on the influence of potential factors such as soil property on the environmental fate of HCHs and DDTs in the PRD has been rare. The present study aimed to investigate the impact of soil physiochemical properties on the distribution patterns and fate of soil HCHs and DDTs on a large spatial scale. Levels of HCHs (sum of α-, ß-, γ- and δ-HCH) and DDTs (sum of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (p,p'-DDT), 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethane (p,p'-DDD), and 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethylene (p,p'-DDE)) in 151 soil samples covering all areas of the PRD and physiochemical parameters related to soil properties including pH, total organic carbon (TOC), total Fe (TFe), DCB-Fe (DFe), amorphous-Fe (AFe), complexed-Fe (CFe), total Mn (TMn), DCB-Mn (DMn), amorphous-Mn (AMn), complexed-Mn (CMn) and cation exchange capacity (CEC) were determined. The residual levels of HCHs and DDTs in soils of the present study, which are mainly controlled by soil TOC and CFe content and varying spatially with land use types, may potentially pose ecological risk to plants and animals. On the other hand, transformation of soil HCHs may be affected by pH and DDT transformation correlated significantly with AFe and CFe. Currently, soil has become an important secondary source of OCPs and the re-emission potential of OCPs in soil was mainly affected by soil OCP concentrations and land use types.