SLR클럽 원문링크 m.slrclub.com/v/hot_article/656080
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IDENTIFICATION OF SEX SPECIFIC
GENES IN AGLAOTHAMNION
CALLOPHYLLIDICOLA
(RHODOPHYTA) USING
DIFFERENTIALLY EXPRESSED
GENE ANALSIS
A R Choi, J B Shim, M Cho and G H Kim
(dofks0704@kongju.ac.kr)
Department of Biology, Kongju National University, Kongju,
Chungnam 314-701, Korea
In red algae, male and female gametophytes have different
morphologies and chemical characteristics so that the
expression of a group of genes is expected during the
sexual development. Our previous proteomic studies
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showed that about 6.1% of proteins are expressed
differentially between male and female gametophytes. Full
cDNA sequence of the proteins, however, was hardly
obtained because of the handicap using degenerated
primers based on the internal sequence of isolated proteins.
In this study, differentially expression genes (DEGs) were
investigated in Aglaothamnion callophyllidicola using annealing control primer (ACP)-based polymerase chain
reaction (PCR) and real-time PCR. Sixty ACP-primers
yielded total of 182 bands and 10 of them were sex
specific; 6 male specific and 4 female specific. The sex
specific bands were eluted and cloned to get partial
sequence of each gene. Validity of the sex specific genes
was examined using real time PCR. Most genes showed 2?5
times difference in quantity between male and female
plants, which was difficult to verify using northern blot
method. A male specific gene was isolated which was
expressed fifty times more in male plants from semiquantitative RT-PCR. Specific primers were designed for 59
and 39 RACE and a full cDNA of 1088 bp was obtained.
Northern blot analysis confirmed that the cDNA was male
specific. BLAST search of the cDNA yielded no significant
match (,30%) to any known genes. Therefore, we named
this noble gene as AMSG1 (A. callophyllidicola Male Sexrelated Gene).
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PHYSIOLOGIC
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