Cloning of vacuolar H+-ATPase subunit c genes from Japanese iris, and functional characterization in yeast
2011
Zhou, Ai-min | Wu, Duo | Che, Dai-di | Liu, Shen-kui | Yang, Chuan-ping | Wang, Jin'gang
Five different isoforms (IrlVHA-c1-c5) of V-ATPase subunit c (VHA-c) were cloned from a Japanese iris (Iris lactea Pall. var. chinensis Fisch. Koidz) cDNA library using degenerate primers PCR and the 5′-RACE technique. The sequence analysis showed the open reading frame (ORF) of the IrlVHA-c1-c5 to be 495 bp, corresponding to a protein of 164 amino acids. Among the five isoforms, IrlVHA-c1 and IrlVHA-c2 are completely homologous. The IrlVHA-c protein is localized at the vacuolar membrane as indicated by a green fluorescent protein (GFP) marker. Its over-expression in yeast could enhance yeast tolerance to NaCl stress. These results show that there are at least five genes encoding different isoforms of IrlVHA-c in Japanese iris and IrlVHA-c is important for the function of V-ATPase.
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