@article{oai:niigata-u.repo.nii.ac.jp:00001569,
 author = {Honjoh, Ken-ichi and Matsumoto, Hiroko and Shimizu, Hideyuki and Ooyama, Kanae and Tanaka, Kageyuki and Oda, Yuichi and Takata, Ryoji and Joh, Toshio and Suga, Koushirou and Miyamoto, Takahisa and Iio, Masayoshi and Hatano, Shoji},
 issue = {8},
 journal = {Bioscience, Biotechnology, and Biochemistry},
 month = {},
 note = {The nucleotide sequence of hiC12, isolated as a cDNA clone of hardening-induced Chlorella (hiC) genes, was identified. The clone encodes a late embryogenesis abundant (LEA) protein having six repeats of a 11-mer amino acid motif, although in a slightly imperfect form. To overexpress the hiC61) and hiC12 genes, their coding regions were PCR amplified and subcloned into a pGEX-1λT vector. The HIC6 and HIC12 proteins were expressed as GST fusion proteins in E. coli, then purified. The two HIC proteins were found to be effective in protecting a freeze-labile enzyme, LDH, against freeze-inactivation. On a molar concentration basis, they were about 3.1×106 times more effective in protecting LDH than sucrose and as effective as BSA. Cryoprotection tests with five kinds of chain-shortened polypeptides, synthesized based on the 11-mer amino acid motif of the HIC6 protein showed that the cryoprotective activity decreased with a decrease in the repeating units of the 11-mer motif. In fact, cryoprotective activities of three kinds of single 11-mer amino acids were very low even at high concentrations. All the results suggested that the sufficiently repeated 11-mer motif is required for the cryoprotective activities of Chlorella LEA proteins.},
 pages = {1656--1663},
 title = {Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27.},
 volume = {64},
 year = {2000}
}