1994 Fiscal Year Final Research Report Summary
Sequence analysis of Rh polypeptide cDNAs and its application to forensic practice
| Project/Area Number |
05807039
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Legal medicine
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KAJII Eiji Jichi Madical School, Dept.of Medicine, Assistant professor, 医学部, 助教授 (40204391)
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| Co-Investigator(Kenkyū-buntansha) |
IKEMOTO Shigenori Jichi Medical School, Dept.of Medicine, Professor, 医学部, 教授 (90048942)
IWAMOTO Sadahiko Jichi Medical School, Dept.of Medicine, Lecturer, 医学部, 講師 (10232711)
TSUCHIDA Shuichi Jichi Medical School, Dept.of Medicine, Lecturer, 医学部, 講師 (20217326)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Rh blood group / Polypeptide / Gene / Cloning / Splicing / Isoform |
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| Research Abstract |
Two Rh polypeptide cDNA have been isolated from the PCR products and tentatively designated RhPI cDNA and RhPII cDNA.Both cDNA clones have an open reading frame composed of 1251 nucleotides. The RhPI cDNA clone shows a single nucleotide substitution with no amino acid substitution compared with the published sequence. The RhPII cDNA clone, on the other hand, differs from the above by 41 nucleotide substitutions with the open reading frame, resulting in 31 amino acid substitutions.
By a systemic analysis of Rh-related mRNA isoforms expressed in reticulocytes, eleven and five truncated isoforms of the RhPI and RhPII cDNAs, respectively, were identified. These isoforms appear to be generated by combinatorial splicing of six RhPI and three RhPII exons. Our results suggest that the Rh-related polypeptide isoforms differing at the C terminus. Multiple RNA splicing pathway are thus operative in the two Rh-related genes even within a single cell lineage of human erythroid cells.
The expression of the Rh gene at the mRNA was analyzed in purified populations of human leukocytes by using RT-PCR followed by Southern blot analysis. The PCR products of the 5^1-terminal region showed a single band as expected in the CD19^+TCR-1^- and CD14^+ cells but doubtfully in the CD13^+CD71^- and CD19^-TCR-1^+ cells. On the other hand, in all these cells, the PCR products of the 3' region exhibited multiple additional bands migrating ahead of the band as expected, which showed distinctly different sets of bands in each cell. The additional bands appeared to consist of RhPI-and RhPII-cDNA splicing isoforms. These results indicated that the expression of the Rh gene is not restricted to human erythroid lineage. Additionally, it was suggested that different transcription initiation sites might be utilized preferentially for Rh gene expression.
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[Publications] Ikemoto, S., Tsuchida, S., Fukui, E., Kajii, E., Iwamoto, S., Kato, K., Akutsu, M., Amemiya, Y.and Miura, Y.: "Further evidence for transformation of genetic markers in recipients after BMT." Adv Foren Haemmogenet. 5. 617-619 (1994)
Description
「研究成果報告書概要(欧文)」より
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