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RANDOM SEQUENCING OF BRASSICA NAPUS cDNAs
Lee, M. H.,Lee, C. M.,Kwak, J. M.,Nam, H. G.,Sohn, U Plant Molecular Biology & Biotechnology Research C 1994 Plant molecular biology and biotechnology research Vol.1994 No.
Single-run cDNA sequencing was performed on 230 cDNA clones of Brassica napus to generate expressed sequence tags (ESTs) and to isolate novel genes from this plant. On average, conventional manual sequencing generated usable sequences approximately 200-250 nucleotides. The ESTs were characterized by comparison with the sequences in the GenBank nucleic acid database and Protein Identification Resource. (PIR) database Of the 213 ESTs generated, 11 ESTs (8.7%) showed significant database-match to the protein coding sequences in the database. Putatively identified ESTs were 21% (50/230) of the total clones. Seventeen clones contained long polyadenylate inserts (7.3%) and 147 clones (64.8%) were unknown genes. Database-matched ESTs include house-keeping genes and genes that are involved in the development, nitrogen fixation, self-incompatibility, pollen specificity, cellular differentiation, photomorphogenesis and photosynthesis in plants. Defense- and stress-related genes and a gene for microbial biogenesis were also detected.
ISOLATION AND CHARACTERIZATION OF A cDNA FROM COLD-INDUCED BRASSICA OLERACEA
Jo, H. R.,Lee, C. M.,Sohn, U-Ik Plant Molecular Biology & Biotechnology Research C 1994 Plant molecular biology and biotechnology research Vol.1994 No.
A cDNA library was constructed from the cold induced B.o leracea leaf and a putative cold induced cDNA, BOCOR, was isolated by the PCR method. A PCR product was cloned into a Bluescript vector. Nucleotide sequence analysis revealed that BOCOR was consisted of 457 bp nucleotides with deduced amino acids of 142 residues. BOCOR showed a high degree of homology to the cold-regulated winter Brassica napus cDNA (BN115). When PCR was carried out using the reverse-transcribe mRNAs from cold-induced and control plants. the 457 bp DNA was amplified only from the cold-induced transcripts. It will be interesting to test if the DNA encodes a cold-regulated protein. Genomic PCR shows that the BOCOR may contain at least one intron in its open reading frame. In an effort to generate cold-resistant transgenic plants, cloning of the cDNA into a plant expression vector is in progress.
MOLECULAR CLONING AND CHARACTERIZATION OF A G PROTEIN β SUBUNIT HOMOLOG, BGB1, FROM BRASSICA NAPUS
Kwak, June-Myoung,Kim, Sun-A,O, Sung-Aeong,Byoun, Chang-Hyoun,Nam, Hong-Gil Plant Molecular Biology & Biotechnology Research C 1994 Plant molecular biology and biotechnology research Vol.1994 No.
We have isolated a cDNA clone, named bgb1, encoding a G protein β subunit homolog from Brassica napus. The predicted 327-residue bgbl, protein (Mr 35,725) shows relatively low homology (20% identity) to mammalian Gβ subunits, but it has a strong homology (60% identity) to RACK1, which is an intracellular receptor for protein kinase C and has a homology to the G protein β subunit (Ron et al., 1994). In addition, bgbl is composed of seven segments of WD-40 repeats that is a well-known characteristic of Gβ subunits. Genomic Southern blot analysis in B. napus and Arabidopsis thaliana revealed that bgbl and its closely related sequences consist of a multigene family in both plants. Approximately 1.4 kilobase transcripts are observed in all organs of B. napus examined by Northern blot analysis. Wound- and touch-treatments do not affect the expression pattern of bgbl in B. napus, but 5 days-dark treatment reduces the bgbl-homologous transcripts in datached leaves of Arabidopsis.
Lim, Chae-Oh,Kim, Min-Gab,Kim, Ho-Yeon,Chung, Woo-Sik,Park, Sung-Han,Hwang, In-Hwan,Jo, Moo-Je Plant Molecular Biology & Biotechnology Research C 1994 Plant molecular biology and biotechnology research Vol.1994 No.
Single-run partial sequencing was performed on randomly selected coda clones from a chinese cabbage flower buds cDNA library. Sequences of seven hundreds sixty nine individual cDNA clones have been compared with sequences in protein database, and three hundreds sixty seven clones (47.7%) showed a varying degree of sequence homology to a variety of known proteins involved in various biological processes. Among the cDNAs with no sequence similarity to known to previously registered other plant ESTs. Most of the EST matched clones also showed sequence similarity with deduced polypeptide sequences of the corresponding ESTs at the amino acid level. Most likely these EST matched cDNAs encode polypeptides conserved in plants. These sequences together with no matched sequences (39.2%) may encode currently unknown flower bud-specific of plant-specific proteins. These Brassica sequences add new EST plants and will be useful to understand genome utilization and genome organization in various tissue within a plant and between different plant species. These EST can also serve as a useful gene resource for basic science in plant molecular biology and for application in agriculture.
PATHOGENIC REACTIONS OF RICE BLAST FUNGI ON DIFFERENTIAL RICE CULTIVARS OF SUSPENSION CELLS
Kang, Kyu-Young,Kim, Seok-Hyeon,Kang, Soo-Woong Plant Molecular Biology & Biotechnology Research C 1994 Plant molecular biology and biotechnology research Vol.1994 No.
The races of rice blast fungi are classified by disease reactions on the differential set of cultivars of rice. These race-cultivar interactions were tested in a single cell level of suspension culture system. The calli were induced from seeds of differential cultivars; Tetep, Kanto51, Nongbaeg, Donghae, Chcueong, and Nagdong, and then transferred into a liquid suspension culture medium. The conidia of rice blast fungus race KJ101 and KJ301 were inoculated directly onto suspension calli and viability were checked and quantified using TTC (2,3,5-triphenyl tetrazolium chloride) staining method. Cell death was observed 3 days after inoculation in susceptible cultivar, Nagdong. The disease reactions turned out to be operated in a single cell level of suspension culture even more elaborately to those of intact plant.