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Molecular cloning of RhD cDNA derived from a gene present in RhD- positive,
but not RhD-negative individuals
MA Arce, ES Thompson, S Wagner, KE Coyne, BA Ferdman and DM Lublin
Department of Pathology, Washington University School of Medicine, St
Louis, MO 63110.
The Rh blood group system plays a major role in immune and nonimmune
hemolytic states. Although an Rh cDNA has been previously cloned, there is
no information on which Rh antigenic protein it encodes. Using polymerase
chain reaction (PCR) amplification, we have identified this original Rh
clone, here designated Rh21, and an additional Rh cDNA clone, Rh13, that is
96% nucleotide- and 92% amino acid-identical to Rh21, with the
substitutions scattered throughout the sequence. A molecular genetic
approach was used to match this Rh clone with an Rh specificity. The mRNA
transcript for Rh13 was present in reticulocytes from RhD-positive
individuals, but was absent from the reticulocytes of RhD-negative
individuals. Using conventional screening of genomic libraries, as well as
PCR cloning, partial genomic clones for these two Rh cDNAs were obtained.
Based on PCR analysis and Southern blots, the Rh21 gene was present in all
individuals, but an intact Rh13 gene was only present in RhD-positive and
not RhD-negative individuals. Thus, by correlating the presence of Rh mRNA
and gene sequences with individual Rh phenotypes, we were able to establish
that the new Rh13 cDNA clone represents the RhD protein.
Volume 82,
Issue 2,
pp. 651-655,
07/15/1993
Copyright © 1993 by The American Society of Hematology
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