Isolation, characterization and expression of a peach nuclear gene encoding a light-harvesting chlorophyll a/b-binding polypeptide of photosystem II
Chung, Kyu Hwan
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https://hdl.handle.net/2142/22623
Description
Title
Isolation, characterization and expression of a peach nuclear gene encoding a light-harvesting chlorophyll a/b-binding polypeptide of photosystem II
Author(s)
Chung, Kyu Hwan
Issue Date
1994
Doctoral Committee Chair(s)
Buetow, Dennis E.
Korban, Schuyler S.
Department of Study
Molecular and Integrative Physiology
Discipline
Molecular and Integrative Physiology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Biology, Botany
Biology, Animal Physiology
Language
eng
Abstract
A nuclear gene encoding a light-harvesting chlorophyll a/b-binding polypeptide of photosystem II has been isolated from a peach genomic library (Prunus persica L. cv. Stark Earliglo) and characterized. The genomic library was constructed and screened with a ($\alpha$-$\sp{32}$P) -labeled pea Lhcb1*1 cDNA probe for positive phage containing Lhcb gene sequences. A phage with an Sstl-restricted peach DNA fragment containing an Lhcb gene sequence was identified. This DNA fragment was subcloned (pAB19) and sequenced completely in both directions. pAB19 contains a complete Lhcb gene coding sequence, designated Lhcb1*Pp1, as well as 5$\sp\prime$ and 3$\sp\prime$ flanking regions. Lhcb1*Pp1 encodes a precursor polypeptide of 267 amino acids composed of a transit peptide of 34 amino acids and a mature polypeptide of 233 amino acids. Transcription initiation and termination sites of Lhcb1*Pp1 mRNA were determined by S1 nuclease mapping. The cap site is located 70 bp upstream from the translation start site and 44 bp downstream from a putative TATA box. Multiple polyadenylation sites are present in the 3$\sp\prime$ flanking region. The seasonal expression of Lhcb1*Pp1 was determined by Northern blot and S1 nuclease protection analyses. Both analyses indicate that the relative level of Lhcb1*Pp1 mRNA per unit total leaf RNA declined to a small extent (27%) from the beginning (June) to the end (September) of the growing season, but decreased by 72% with senescence in October. Also, S1 nuclease protection analyses with 5$\sp\prime$ and 3$\sp\prime$ gene probes showed that Lhcb1*Pp1 is expressed in three varieties of P. persica and in light-grown but its expression is undetectable in dark-grown leaves and stems of the Evergreen variety.
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