This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone （r-fGH） was expressed by Escherichia coli in form of inclusion body or as ...This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone （r-fGH） was expressed by Escherichia coli in form of inclusion body or as soluble protein under different inducing conditions. The inclusion body was renatured using two recovery methods, i.e., dilution and dialysis. Thereafter, the refolded protein was purified by Glutathione Sepharase 4B affinity chromatography and r-fGH was obtained by cleavage of thrombin. For soluble products, r-fGH was directly purified from the lysates by Glutathione Sepharase 4B affinity chromatography. ELISA-receptor assay demonstrated that despite its low receptor binding activity, the r-fGH purified from refolded inclusion body had a higher yield （2.605 mg L^-1） than that from soluble protein （1.964 mg L^-l）. Of the tested recovery methods, addition of renaturing buffer （pH 8.5） into denatured inclusion body yielded the best recovery rate （17.9%）. This work provided an optimized purification method for high recovery of r-fGH, thus contributing to the application of r-fGH to aquaculture.展开更多
This experiment was conducted to optimize the culture conditions to induce calli from chilli pepper （Capsicum annuum L.） local cultivar. The mature seeds were surface sterilized for one min with 70% Ethanol followed...This experiment was conducted to optimize the culture conditions to induce calli from chilli pepper （Capsicum annuum L.） local cultivar. The mature seeds were surface sterilized for one min with 70% Ethanol followed by 20 min with （0%, 2%, 4% or 6%） NaOCI and were germinated on MS medium with 2 mg/L GA3. Seedlings and mature fruits were used as explants source. The placenta, pericarp, hypocotyls, cotyledonal leaves, shoot tips and roots were cultured on MS media supplemented with Kinetin （0.0 mg/L, 0.5 mg/L, 1.0 mg/L, 2.0 mg/L） and IAA （0.0 mg/L, 1.0 mg/L, 2.0 mg/L） in different combinations or NAA or 2,4-D （0.0 mg/L, 1.0 mg/L, 2.0 mg/L, 4.0 mg/L）. Callus fresh weight was recorded after 4 weeks in culture. The results showed that the best sterilizing method was with 70% Ethanol followed by 20 mints with （4% or 6%） NaOCI, however 6% NaOCI reduced seed＇s viability. Callus was induced from all explants cultured on MS media supplemented with IAA and Kinetin except the placenta and the pericarp. The results showed that the hypocotyls surpass all other explants in the mean callus fresh weight which was 160.58 mg compared with 147.81 mg, 134.95 mg, and 122.33 mg for cotyledonal leaves, shoot tips and roots respectively. Moreover the analysis of the interaction between the growth regulators and the explants showed that 2 mg/L IAA and Kinetin had significant effect on callus mean fresh weight which was （309.74, 339.14, 358.48, and 284.64） mg for the shoot tips, cotyledonal leaves, hypocotyls and roots, respectively. On the other hand, 2 mg/L 2,4-D or NAA was the best concentration for callus induction from the placenta and the pericarp. The pericarp gave a mean fresh weight of 276.90 mg in the presence of 2,4-D compared with 253.60 mg for the placenta. Moreover the pericarps gave significantly higher fresh weight than the placenta with an average of 210.3mg and 184.9 mg respectively in the presence of 2 mg/L NAA. In conclusion the best sterilization method of chilli pepper seeds is by 70% ethanol for one minute followed by 20 min in 4% （NaOCI）. The best explants for callus induction only is the Hypocotyls grown on MS medium supplemented with 2 mg/L oflAA and Kinetin under the conditions of the current experiment.展开更多
The relationships among the proportions of the neon dual isotopes ratio, scale factor corrections (SFCs), light intensities, environmental magnetic field and magnetic zero drift are discussed in detail by numerical si...The relationships among the proportions of the neon dual isotopes ratio, scale factor corrections (SFCs), light intensities, environmental magnetic field and magnetic zero drift are discussed in detail by numerical simulations. The results show that the unification of the optimal operating point (OP) and the frequency stabilization operating point (FSP) is achievable by adjusting the proportions of neon dual isotopes accurately and tuning the cavity length with frequency stabilization system exactly. In that case, the left-rotation and right-rotation gyros can obtain the same SFC, which can decrease the magnetic sensitivity of the laser gyro efficiently. The Zeeman effect zero drift and the Faraday bias zero drift are both reduced by two orders of magnitude, while the magnetic shielding requirement of laser tube is decreased by 1-2 orders of magnitude.展开更多
基金supported by the National Natural Science Foundation of China (No.30901111)the China Agriculture Research System (CARS-50)the key Project of Chinese Ministry of Education (No.108083)
文摘This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone （r-fGH） was expressed by Escherichia coli in form of inclusion body or as soluble protein under different inducing conditions. The inclusion body was renatured using two recovery methods, i.e., dilution and dialysis. Thereafter, the refolded protein was purified by Glutathione Sepharase 4B affinity chromatography and r-fGH was obtained by cleavage of thrombin. For soluble products, r-fGH was directly purified from the lysates by Glutathione Sepharase 4B affinity chromatography. ELISA-receptor assay demonstrated that despite its low receptor binding activity, the r-fGH purified from refolded inclusion body had a higher yield （2.605 mg L^-1） than that from soluble protein （1.964 mg L^-l）. Of the tested recovery methods, addition of renaturing buffer （pH 8.5） into denatured inclusion body yielded the best recovery rate （17.9%）. This work provided an optimized purification method for high recovery of r-fGH, thus contributing to the application of r-fGH to aquaculture.
文摘This experiment was conducted to optimize the culture conditions to induce calli from chilli pepper （Capsicum annuum L.） local cultivar. The mature seeds were surface sterilized for one min with 70% Ethanol followed by 20 min with （0%, 2%, 4% or 6%） NaOCI and were germinated on MS medium with 2 mg/L GA3. Seedlings and mature fruits were used as explants source. The placenta, pericarp, hypocotyls, cotyledonal leaves, shoot tips and roots were cultured on MS media supplemented with Kinetin （0.0 mg/L, 0.5 mg/L, 1.0 mg/L, 2.0 mg/L） and IAA （0.0 mg/L, 1.0 mg/L, 2.0 mg/L） in different combinations or NAA or 2,4-D （0.0 mg/L, 1.0 mg/L, 2.0 mg/L, 4.0 mg/L）. Callus fresh weight was recorded after 4 weeks in culture. The results showed that the best sterilizing method was with 70% Ethanol followed by 20 mints with （4% or 6%） NaOCI, however 6% NaOCI reduced seed＇s viability. Callus was induced from all explants cultured on MS media supplemented with IAA and Kinetin except the placenta and the pericarp. The results showed that the hypocotyls surpass all other explants in the mean callus fresh weight which was 160.58 mg compared with 147.81 mg, 134.95 mg, and 122.33 mg for cotyledonal leaves, shoot tips and roots respectively. Moreover the analysis of the interaction between the growth regulators and the explants showed that 2 mg/L IAA and Kinetin had significant effect on callus mean fresh weight which was （309.74, 339.14, 358.48, and 284.64） mg for the shoot tips, cotyledonal leaves, hypocotyls and roots, respectively. On the other hand, 2 mg/L 2,4-D or NAA was the best concentration for callus induction from the placenta and the pericarp. The pericarp gave a mean fresh weight of 276.90 mg in the presence of 2,4-D compared with 253.60 mg for the placenta. Moreover the pericarps gave significantly higher fresh weight than the placenta with an average of 210.3mg and 184.9 mg respectively in the presence of 2 mg/L NAA. In conclusion the best sterilization method of chilli pepper seeds is by 70% ethanol for one minute followed by 20 min in 4% （NaOCI）. The best explants for callus induction only is the Hypocotyls grown on MS medium supplemented with 2 mg/L oflAA and Kinetin under the conditions of the current experiment.
基金supported by the Science Foundation of Aeronautics of China (No.20090853014)
文摘The relationships among the proportions of the neon dual isotopes ratio, scale factor corrections (SFCs), light intensities, environmental magnetic field and magnetic zero drift are discussed in detail by numerical simulations. The results show that the unification of the optimal operating point (OP) and the frequency stabilization operating point (FSP) is achievable by adjusting the proportions of neon dual isotopes accurately and tuning the cavity length with frequency stabilization system exactly. In that case, the left-rotation and right-rotation gyros can obtain the same SFC, which can decrease the magnetic sensitivity of the laser gyro efficiently. The Zeeman effect zero drift and the Faraday bias zero drift are both reduced by two orders of magnitude, while the magnetic shielding requirement of laser tube is decreased by 1-2 orders of magnitude.
