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Biochemistry, Biophysics, and Structural Biology Commons

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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

An Expanded Toolkit For Gene Tagging Based On Mimic And Scarless Crispr Tagging In, David Li-Kroeger, Oguz Kanca, Pei-Tseng Lee, Sierra Cowan, Michael T Lee, Manish Jaiswal, Jose Luis Salazar, Yuchun He, Zhongyuan Zuo, Hugo J Bellen Aug 2018

An Expanded Toolkit For Gene Tagging Based On Mimic And Scarless Crispr Tagging In, David Li-Kroeger, Oguz Kanca, Pei-Tseng Lee, Sierra Cowan, Michael T Lee, Manish Jaiswal, Jose Luis Salazar, Yuchun He, Zhongyuan Zuo, Hugo J Bellen

Faculty Publications

We generated two new genetic tools to efficiently tag genes in Drosophila. The first, Double Header (DH) utilizes intronic MiMIC/CRIMIC insertions to generate artificial exons for GFP mediated protein trapping or T2A-GAL4 gene trapping in vivo based on Cre recombinase to avoid embryo injections. DH significantly increases integration efficiency compared to previous strategies and faithfully reports the expression pattern of genes and proteins. The second technique targets genes lacking coding introns using a two-step cassette exchange. First, we replace the endogenous gene with an excisable compact dominant marker using CRISPR making a null allele. Second, the insertion is replaced …


Analyzing The Effects Of Site-Directed Mutagenesis On The Structure Of Coxsackievirus B3 Genomic Rna, Erin Dimon May 2018

Analyzing The Effects Of Site-Directed Mutagenesis On The Structure Of Coxsackievirus B3 Genomic Rna, Erin Dimon

Theses/Capstones/Creative Projects

Coxsackievirus B3 (CVB3) is known to cause myocarditis and pancreatitis in humans. The virus has a single stranded RNA genome that codes for 11 different proteins. CVB3 is found to have two serotypes: 28 (virulent and disease causing) and GA (avirulent and not disease causing). Like other members of the Picornaviridae family, CVB3 utilizes the genomic 5’ untranslated region (5’ UTR) to initiate viral replication mechanisms through interactions with host protein factors. Structural variations of the CVB3 5’ UTR are hypothesized to influence the success of such interactions and consequentially determine viral virulence. The aim of this project was to …


A Multisession, Undergraduate Molecular Biology Lab Experiment Using Green Fluorescent Protein Including Subcloning And Color Changing Mutagenesis, Nathan S. Winter Apr 2018

A Multisession, Undergraduate Molecular Biology Lab Experiment Using Green Fluorescent Protein Including Subcloning And Color Changing Mutagenesis, Nathan S. Winter

Chemistry Faculty Publications

This paper describes a series of experiments involving handling and manipulating the DNA coding for Green Fluorescent Protein (GFP) including the subcloning of this gene, and mutating the DNA so that Cyan Fluorescent Protein (CFP) or Blue Fluorescent protein (BFP) are expressed. The primers needed for the PCR based subcloning of GFP are presented, as are those needed to mutate the GFP to either CFP or BFP.