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Nucleic Acids Research, 1989, Vol. 17, No. 22 9267-9277
© 1989

MOLECULAR BIOLOGY

Characterization of amplified intracisternal A-particle elements encoding integrase

Kira K. Lueders*, Zehava Grossman+ and Joseph W. Fewell

Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health Bethesda, MD 20892, USA

*To whom correspondence should be addressed

Received July 17, 1989. Revised October 2, 1989. Accepted October 2, 1989.

Type IIB intracisternal A-particle (IAP) elements have undergone marked amplification and transposition in the genomic DNA of some mouse myelomas. We have made a cDNA library from one such myeloma, MOPC 315, to determine whether some property of the elements themselves has a role in this process. Sequencing of several type IIB cDNAs and one genomic type HE IAP element has shown that they are nearly identical (>99%) and contain 2 open reading frames (ORFs). ORF2 is capable of encoding the IAP integrase, an enzyme which catalyzes integration of proviral DNA into the genome. An antiserum to a synthetic peptide based on the IAP integrase gene sequence reacted with ORF2 product expressed in bacteria as a fusion protein, and detected a 47 kDa protein, predicted from the size of ORF2, in myeloma cell fractions by Western blotting.

+Present address: Institute of Hematology, Chaim Sheba Medical Center, Hashomer, Israel


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