2 research outputs found
Recombinant expression of marine shrimp lysozyme in Escherichia coli
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
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