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. 2016 Feb 24;7:176. doi: 10.3389/fpls.2016.00176

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Copyright © 2016 Sabila, Kundu, Smalls and Ullah.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Stable expression of bait and prey constructs. (A) PCR amplification of bait construct using RACK1A and bait specific (lane 1- AA; lane 2- YY) or RACK1A and prey specific (lane 3- AA and lane 4- YY) primers. The empty vector containing EE cells did not show any amplification from either primer pairs (lane 5 and lane 6). No template control (lane 7) did not amplify any band. (B) Yeast cells maintaining both the bait and prey constructs [pMKZ-RACK1A:NUI-RACK1A (abbreviated as AA) and pMKZ- RACK1A-Y248F:NUI-RACK1A-Y248F (abbreviated as YY)] are able to grow of media lacking both the selection markers (minus histidine and tryptophan)- confirming stable expression of the constructs in the AA and YY yeast cells. (C) Serial dilution of the individual prey constructs (pMKZ-RACK1A (left upper) and pMKZ-RACK1A-Y248F (left lower) are able to grow on the SD-H selection plates while the individual bait constructs (NUI-RACK1A (right upper) and NUI-RACK1A-Y248F (right lower) are able to grow on the bait specific selection plates (SD-T).