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Biomedical subjects

H Sawaki

Publications and source records attributed to H Sawaki.

4 recordsLinked to original sources
PubMed2001

Expressed genes in the extraradical hyphae of an arbuscular mycorrhizal fungus, Glomus intraradices, in the symbiotic phase.

To collect extraradical hyphae of arbuscular mycorrhizal (AM) fungi for RNA isolation, a PVDF membrane was laid on the hyphal compartment of a two-compartment culture system of transformed carrot hairy roots and Glomus intraradices. Extraradical hyphae free from host tissue were easily collected, and their RNA was rapidly extracted with a modified acid guanidinium thiocyanate-phenol-chloroform method. A 3'-RACE (rapid amplification of cDNA ends) of a known gene indicated that this protocol enabled the isolation of mRNA molecules as small as 2.3 kb. The cDNA libraries of an AM fungus from the aseptic extraradical hyphae in a symbiotic state were constructed for the first time. Three-fourth of 150 ESTs (expressed sequence tags) indicated low or no similarities to known sequences from other organisms.

DNA, Complementary↗
PubMed1998

Promoters of the phycocyanin gene clusters of the cyanobacterium Synechococcus sp. strain PCC 7942.

The cyanobacterium Synechococcus sp. strain PCC 7942 has duplicated phycocyanin subunit gene clusters cpcB1A1 and cpcB2A2, which are identical to each other and to those of Synechococcus sp. strain PCC 6301 (Anacystis nidulans). Nucleotide sequences of the 428 and 286 bases of the 5' non-coding regions of the cpcB1A1 and cpcB2A2 clusters, respectively, of strain PCC 7942 were identical to those of strain PCC 6301. As in strain PCC 6301, cpcB1A1 yielded two major transcripts of 1.4 and 1.3 kb and cpcB2A2 yielded a single transcript of 1.3 kb in strain PCC 7942. Thus, the structure and expression of cpcBA gene clusters in the two strains are essentially the same. Using bacterial luciferase encoded by luxAB as a reporter, cpcB1A1 was shown to have two promoters corresponding to the two major transcripts. Luminescence from the Synechococcus reporter strains carrying the fusions of the cpcBA promoters to luxAB showed circadian oscillation. Similar to the promoter of psbA1 encoding the D1 protein of PSII, the two cpcB1A1 promoters and the cpcB2A2 promoter showed the peak of activity at the end of the subjective day and the trough at the end of the subjective night.

Base Sequence↗
PubMed1992

Cloning and sequence analysis of cDNA encoding aspartate aminotransferase isozymes from Panicum miliaceum L., a C4 plant.

The cytosolic and mitochondrial isozymes of aspartate aminotransferase (AspAT) function in the C4 dicarboxylate cycle of photosynthesis. We constructed a cDNA library from leaf tissues of Panicum miliaceum, an NAD-malic-enzyme-type C4 plant and screened the library for AspAT isozymes. A full-length cDNA clone for cytosolic AspAT was isolated. This clone contains an open reading frame that encodes 409 amino acids. We also isolated two cDNA clones for different precursors of mitochondrial AspAT. Comparing these two sequences in the coding regions, we found 12 amino acid substitutions out of 28 base substitutions. The encoded amino acid sequences predict that mitochondrial AspAT are synthesized as precursor proteins of 428 amino acid residues, which each consist of a mature enzyme of 400 amino acid residues and a 28-amino-acid presequence. This prediction coincides with the observation that the in vitro translation product of the mRNA for mitochondrial AspAT was substantially larger than the mature form. A comparison of the amino acid sequences of the AspAT isozymes from P. miliaceum with the published sequences for the enzymes from various animals and microorganisms reveals that functionally and/or structurally important residues are almost entirely conserved in all AspAT species.

Amino Acid Sequence↗