Fast and Efficient Expression of Multiple Proteins in Avian Embryos Using mRNA Electroporation

10.3791/59664 ◽  
2019 ◽  
Author(s):  
Martin Tran ◽  
Mohit Dave ◽  
Rusty Lansford
Keyword(s):  
Avian Embryos ◽  
Efficient Expression ◽  
Mrna Electroporation

scholarly journals Novel molecular biological tools for the efficient expression of fungal lytic polysaccharide monooxygenases in Pichia pastoris

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Lukas Rieder ◽  
Katharina Ebner ◽  
Anton Glieder ◽  
Morten Sørlie
Keyword(s):  
Pichia Pastoris ◽  
Biomass Conversion ◽  
Single Step ◽  
Additional Advantage ◽  
Lytic Polysaccharide Monooxygenases ◽  
Expression Host ◽  
Shake Flask Cultivation ◽  
Polysaccharide Monooxygenases ◽  
Efficient Expression ◽  
High Yields

Abstract Background Lytic polysaccharide monooxygenases (LPMOs) are attracting large attention due their ability to degrade recalcitrant polysaccharides in biomass conversion and to perform powerful redox chemistry. Results We have established a universal Pichia pastoris platform for the expression of fungal LPMOs using state-of-the-art recombination cloning and modern molecular biological tools to achieve high yields from shake-flask cultivation and simple tag-less single-step purification. Yields are very favorable with up to 42 mg per liter medium for four different LPMOs spanning three different families. Moreover, we report for the first time of a yeast-originating signal peptide from the dolichyl-diphosphooligosaccharide-protein glycosyltransferase subunit 1 (OST1) form S. cerevisiae efficiently secreting and successfully processes the N-terminus of LPMOs yielding in fully functional enzymes. Conclusion The work demonstrates that the industrially most relevant expression host P. pastoris can be used to express fungal LPMOs from different families in high yields and inherent purity. The presented protocols are standardized and require little equipment with an additional advantage with short cultivation periods.

scholarly journals Evaluating the Observed Log-Likelihood Function in Two-Level Structural Equation Modeling with Missing Data: From Formulas to R Code

Psych ◽  
2021 ◽  
Vol 3 (2) ◽  
pp. 197-232
Author(s):  
Yves Rosseel
Keyword(s):  
Structural Equation ◽  
Structural Equation Models ◽  
Likelihood Function ◽  
Likelihood Estimation ◽  
Missing At Random ◽  
Equation Modeling ◽  
Computationally Efficient ◽  
Log Likelihood ◽  
Efficient Expression ◽  
Special Case

This paper discusses maximum likelihood estimation for two-level structural equation models when data are missing at random at both levels. Building on existing literature, a computationally efficient expression is derived to evaluate the observed log-likelihood. Unlike previous work, the expression is valid for the special case where the model implied variance–covariance matrix at the between level is singular. Next, the log-likelihood function is translated to R code. A sequence of R scripts is presented, starting from a naive implementation and ending at the final implementation as found in the lavaan package. Along the way, various computational tips and tricks are given.

scholarly journals Viral DNA synthesis is required for the efficient expression of specific herpes simplex virus type 1 mRNA species

Virology ◽  
1980 ◽  
Vol 101 (1) ◽  
pp. 10-24 ◽  
Author(s):  
Louis E. Holland ◽  
Kevin P. Anderson ◽  
Charles Shipman ◽  
Edward K. Wagner
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Synthesis ◽  
Virus Type ◽  
Herpes Simplex Virus Type ◽  
Viral Dna ◽  
Mrna Species ◽  
Efficient Expression ◽  
Simplex Virus

scholarly journals FISHing for chick genes: Triple-label whole-mount fluorescence in situ hybridization detects simultaneous and overlapping gene expression in avian embryos

2004 ◽  
Vol 229 (3) ◽  
pp. 651-657 ◽  
Author(s):  
Nathaniel Denkers ◽  
Pilar García-Villalba ◽  
Christopher K. Rodesch ◽  
Kandice R. Nielson ◽  
Teri Jo Mauch
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Avian Embryos ◽  
Overlapping Gene ◽  
Whole Mount
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