|Budget Amount *help
¥2,000,000 (Direct Cost : ¥2,000,000)
Fiscal Year 1995 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1994 : ¥1,600,000 (Direct Cost : ¥1,600,000)
A processed gene was identified in the intergenic alpha-globin gene region in the macaques. This sequence was 575 bp in length and included initiation and termination codons, signal sequence for polyA and some length of A rich region at 3' terminal. It was flanked by a short direct repeat. Since the open reading frame of this gene encoded a protein consisting 117 amino acids, we named this processed gene P117. The parental gene for P117 was expressed in the liver, brain, kidney and lung. All of primates showd at least one positive band in Southern blot analysis. The processed gene was inserted among primates widely from prosimians to hominoids. The insertion of the processed P117 gene was examined in 385 heads macaques. The genetic frequency were variable in macaques, for example, 0.642 in crab-eating macaques from eastern part in Thailand, 0.550 in rhesus macaques from India but 0% in Japanese macaques and six Sulawesi macaques.
The nucleotide sequence of the processed P117 amplified by PCR method from seven species of primates was determined. The philogenic tree deduced from the sequences were different from the philogenic tree deduced from other genomic DNA sequences in the common marmosett and in hamadrias baboon. To determine the insertion period of P117, more sequences have to be determined and be compared with the parental P117 gene.
It is necessary to obtain P117 protein to make antibody for the knowledge of function in the several organs. The processed P117 gene was recombined into pGEX 5X-3 vector and transformed into E.coli JM109. The fusion protein with the glutathion-s-transferase was included into the inclusion body. It was disolved in the 5M urea/1XPBS solution. Now the method of separation of P117 prtein from other proteins.