Generating HA-tagged α-catenin Mutant Constructs

Expression vectors containing the HA-tagged α-catenin mutant were previously described (Ozawa, 1998 (link); Matsubara and Ozawa, 2001 (link)). A CAG vector containing HA-tagged α-catenin (pC-αHA) (Taniguchi et al., 2005 (link)) was digested with NotI and EcoRV, and the fragment containing full-length α-catenin cDNA was cloned into the NotI/EcoRV site of pU-DNCT (Ozawa and Kobayashi, 2014 (link)), a derivative of pUHD10-3 (Gossen and Bujard, 1992 (link)), yielding pTRE-α-cateninFLAG. pCAGGShyg, which confers hygromycin resistance, and the pCAG/bsr-7 vector, which confers blasticidin resistance gene, were described previously (Ozawa and Kobayashi, 2014 (link); Ozawa, 2015 (link)). pZeoSV2(+), which contains the zeocin-resistance gene, and the gRNA expression vector containing the α-1,3-galactosyltransferase gene were provided by Masahiro Sato (Kagoshima University). pTRE-GFP–NMHCIIA (Addgene plasmid # 10844) were gifts from Robert Adelstein (Wei and Adelstein, 2000 (link)).

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, & Ozawa M. (2018). Nonmuscle myosin IIA is involved in recruitment of apical junction components through activation of α-catenin. Biology Open, 7(5), bio031369.

Publication 2018

pTRE-GFP–NMHCIIA

Catenin Cdna Galactosyltransferase Gene Gifts Hygromycin Plasmid Vector Zeocin

Corresponding Organization : Kagoshima University

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Variable analysis

independent variables

HA-tagged α-catenin mutant expression vectors
PTRE-α-cateninFLAG vector
PCAGGShyg vector (confers hygromycin resistance)
PCAG/bsr-7 vector (confers blasticidin resistance)
PZeoSV2(+) vector (contains zeocin-resistance gene)
GRNA expression vector containing the α-1,3-galactosyltransferase gene
PTRE-GFP–NMHCIIA vector

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

PCAGGShyg vector (confers hygromycin resistance)
PCAG/bsr-7 vector (confers blasticidin resistance)
PZeoSV2(+) vector (contains zeocin-resistance gene)

negative controls

Not explicitly mentioned

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