2 research outputs found

    Recombinant expression of marine shrimp lysozyme in Escherichia coli

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    Shrimp Lysozyme (Lyz) is a key component of the antibacterial response as part of the innate defense in Crustacea; however, it has not been possible to purify this protein because of the very low amount present in the shrimp blood cells (hemocytes). In an effort to produce enough protein to study its function and biochemical properties we have overexpressed Lysozyme from marine shrimp ( Penaeus vannamei ) in E. coli. A bacterial protein expression system based on the T7 polymerase promoter was used. Although Lyz was produced as insoluble protein in inclusion bodies, its refolding led to an active protein with a yield of ~10%. Details of the protein recombinant expression techniques applied to this shrimp protein are presented

    Total RNA quality of lyophilized and cryopreserved dormant grapevine buds

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    Background: Plant tissues must be preserved in their collection state, especially for genome-wide expression profile studies. Lyophilization is a feasible, affordable tool when fresh tissues cannot be shipped at ultralow temperatures from their origin to the place of analysis. In this study, the total RNA quality of dormant grapevine buds ( Vitis vinifera L. cv. \u2018Flame Seedless\u2019) of freeze-dried samples stored at room temperature conditions was evaluated and compared to that of cryopreserved (-80\ub0C) grapevine buds. Results: Good yield and quality of RNA were obtained from freeze-dried dormant buds stored at room temperature for 0, 3 and 6 weeks after they were lyophilized. Further experiments confirmed that the extracted total RNA could be used for actin and \u3b2-tubulin PCR gene amplification. Conclusion: High-quality RNA that is useful for downstream applications was obtained from freeze-dried dormant grapevine bud tissue, similarly to the RNA obtained from cryopreserved dormant grapevine buds
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