cDNA cloning and functional expression of the α-d-galactose-binding lectin frutalin in escherichia coli

cDNA clones encoding frutalin, the α-d-galactose-binding lectin expressed in breadfruit seeds (Artocarpus incisa), were isolated and sequenced. The deduced amino acid sequences indicated that frutalin may be encoded by a family of genes. The NCBI database searches revealed that the frutalin sequence is highly homologous with jacalin and mornigaG sequences. Frutalin cDNA was re-amplified and cloned into the commercial expression vector pET-25b(+) for frutalin production in Escherichia coli. An experimental factorial design was employed to maximise the soluble expression of the recombinant lectin. The results indicated that temperature, time of induction, concentration of IPTG and the interaction between the concentration of IPTG and the time of induction had the most significant effects on the soluble expression level of recombinant frutalin. The optimal culture conditions were as follows: induction with 1 mM IPTG at 22°C for 20 h, yielding 16 mg/l of soluble recombinant frutalin. SDS-PAGE and Western blot analysis revealed that recombinant frutalin was successfully expressed by bacteria with the expected molecular weight (17 kDa). These analyses also showed that recombinant frutalin was mainly produced as insoluble protein. Recombinant frutalin produced by bacteria revealed agglutination properties and carbohydrate-binding specificity similar to the native breadfruit lectin.Fundação para a Ciência e a Tecnologia (FCT

New 40Ar/39Ar age constraints on the Late Palaeozoic tectonic evolution of the western Tianshan (Xinjiang, northwestern China), with emphasis on Permian fluid ingress

Laser-probe dating of mylonite whole-rock samples from the North Tianshan - Main Tianshan fault zone that cross-cuts the North Tianshan domain's southern margin yielded 40Ar/39Ar spectra with 255-285 Ma ages. Biotite from an undeformed, Early Carboniferous granite, which cuts the steep mylonitic foliation in the Proterozoic basement of the Yili arcs's southern margin, gave a 263.4 ± 0.6 Ma plateau age (1σ). Pre-Carboniferous metasediments overlying this basement yielded plateau ages (1σ) of 253.3 ± 0.3 (muscovite) and 252.3 ± 0.3 (biotite) Ma. The Permian ages of mylonites date movement on these ductile, dextral strike-slip shear zones, whereas the mica ages are interpreted by recrystallisation as a result of fluid flow around such transcurrent faults. We propose that the Tianshan's Permian syn-tectonic bimodal magmatism was created in a non-plume-related Yellowstone-like extensional - transtensional tectonic regime. Gold mineralisation, tracing aqueous flow in the crust, peaked in Permian time and continued locally into the Triassic. The picture is emerging that a convective fluid system partly driven by magmatic heat, existed in a strongly fractured and weakened crust with an elevated heat flow, leading to regional-scale isotope resetting. We suggest that surprisingly young isotopic ages in the literature for early orogenic (ultra)high-pressure metamorphism are similarly due to fluid-mediated recrystallisation. © Springer-Verlag 2008.link_to_subscribed_fulltex