Baculoviral Expression of Huntingtin Mutants

Expression of full-length Q23-, Q46-, and Q78-huntingtin from our pALHDQ23, pALHDQ46, and pALHDQ78 constructs was done using the Baculovirus Expression system (Invitrogen).⁸ (link) Three lysine aspartic acid (KD) mutant huntingtin constructs (K2449D, K2932D/K2934D, K2449D/K2932D/K2934D) were generated by submission of the DNA sequences to Genscript (Piscataway, NJ, USA), which provided synthesized DNA in the pFastBac1 vector using SalI/SacII restriction digest and standard molecular biology techniques. The preparation and purification of full-length huntingtin was carried out as previously described.⁸ (link)

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Shin B., Jung R., Oh H., Owens G.E., Lee H., Kwak S., Lee R., Cotman S.L., Lee J.M., MacDonald M.E., Song J.J., Vijayvargia R, & Seong I.S. (2018). Novel DNA Aptamers that Bind to Mutant Huntingtin and Modify Its Activity. Molecular Therapy. Nucleic Acids, 11, 416-428.

Publication 2018

Baculovirus Expression system

Baculovirus Dna sequences Huntingtin Lysine acid Vector

Corresponding Organization : Harvard University

Other organizations : California Institute of Technology, Korea Advanced Institute of Science and Technology, CHDI Foundation

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Variable analysis

independent variables

Expression of full-length Q23-huntingtin
Expression of full-length Q46-huntingtin
Expression of full-length Q78-huntingtin
Expression of three lysine aspartic acid (KD) mutant huntingtin constructs (K2449D, K2932D/K2934D, K2449D/K2932D/K2934D)

dependent variables

Not explicitly mentioned

control variables

Use of the Baculovirus Expression system (Invitrogen) for expression of full-length huntingtin
Preparation and purification of full-length huntingtin as previously described

controls

Positive control: Expression of full-length Q23-huntingtin, Q46-huntingtin, and Q78-huntingtin
Negative control: Not explicitly mentioned

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