TY - JOUR
T1 - Cloning and expression of cDNAs encoding plant V-ATPase subunits in the corresponding yeast null mutants
AU - Aviezer-Hagai, Keren
AU - Nelson, Hannah
AU - Nelson, Nathan
N1 - Funding Information:
This work was supported by a grant from the United States–Israel Binational Agricultural Research and Development Fund. We thank Vered Padler-Karavani for providing the VMA11 and VPH1/STV1 yeast null mutants, Natalie Perzov for providing the VMA10 yeast null mutant, Tamar R. Grossman for her help in the preparation of this manuscript and Nathan and Rama Kaminer for providing the lemon fruits.
PY - 2000/8/15
Y1 - 2000/8/15
N2 - Complementation of yeast null mutants is widely used for cloning of homologous genes from heterologous sources. We have used this method to clone the relevant V-ATPase genes from lemon fruit and Arabidopsis thaliana cDNA libraries. The pH levels are very different in the vacuoles of the lemon fruit and the A. thaliana, yet both are the result of the activity of the same enzyme complex, namely the V-ATPase. In order to investigate the mechanism that enables the enzyme to maintain such differences in pH values, we have compared the subunit composition of the V-ATPase complex from both sources. Towards this end, we have constructed a cDNA library from lemon fruit and cloned it into a similar shuttle vector to the one of the A. thaliana cDNA library, which is commercially available. In this work, we report the cloning and expression of VMA10 from both sources, two isoforms of the lemon proteolipid (VMA3) and the lemon homologue of yeast VPH1/STV1 subunit, LEMAC.
AB - Complementation of yeast null mutants is widely used for cloning of homologous genes from heterologous sources. We have used this method to clone the relevant V-ATPase genes from lemon fruit and Arabidopsis thaliana cDNA libraries. The pH levels are very different in the vacuoles of the lemon fruit and the A. thaliana, yet both are the result of the activity of the same enzyme complex, namely the V-ATPase. In order to investigate the mechanism that enables the enzyme to maintain such differences in pH values, we have compared the subunit composition of the V-ATPase complex from both sources. Towards this end, we have constructed a cDNA library from lemon fruit and cloned it into a similar shuttle vector to the one of the A. thaliana cDNA library, which is commercially available. In this work, we report the cloning and expression of VMA10 from both sources, two isoforms of the lemon proteolipid (VMA3) and the lemon homologue of yeast VPH1/STV1 subunit, LEMAC.
KW - Arabidopsis thaliana
KW - Expression cloning
KW - Lemon fruit
KW - V-ATPase
KW - cDNA library
UR - http://www.scopus.com/inward/record.url?scp=0034663501&partnerID=8YFLogxK
U2 - 10.1016/S0005-2728(00)00188-2
DO - 10.1016/S0005-2728(00)00188-2
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AN - SCOPUS:0034663501
SN - 0005-2728
VL - 1459
SP - 489
EP - 498
JO - Biochimica et Biophysica Acta - Bioenergetics
JF - Biochimica et Biophysica Acta - Bioenergetics
IS - 2-3
ER -