cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN)
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cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN). / Young, M F; Kerr, J M; Termine, J D; Wewer, U M; Wang, M G; McBride, O W; Fisher, L W.
In: Genomics, Vol. 7, No. 4, 01.08.1990, p. 491-502.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN)
AU - Young, M F
AU - Kerr, J M
AU - Termine, J D
AU - Wewer, U M
AU - Wang, M G
AU - McBride, O W
AU - Fisher, L W
PY - 1990/8/1
Y1 - 1990/8/1
N2 - A human osteopontin (OP) cDNA was isolated from a library made from primary cultures of human bone cells. The distribution of osteopontin mRNA in human tissues was investigated by Northern analysis and showed that the human message was predominant in cultures of bone cells and in decidua cells isolated at 6-12 weeks of gestation. Immunohistochemical analysis confirmed that OP expression is high in decidua cells as well as in the endometrial glands of a non-pregnant secretory-phase human uterus. Two variants of the OP message were evident on the basis of DNA sequencing and polymerase chain reaction amplification of bone and decidua cell mRNA. The peptides potentially translated by the variant messages differ by the presence (OP1b) or absence (OP1a) of 14 amino acids at residue 58 of the molecule. The deduced human protein sequence shows a conservation between species in the position of the Arg-Gly-Asp (RGD) cell attachment site. Chromosomal mapping of the osteopontin gene (OPN) using human-rodent cell hybrids demonstrated a location on chromosome 4 in the human genome. In situ hybridization of metaphase chromosomes using radiolabeled OP1a as a probe indicated that the gene is located on a region of 4q that is near the centromere. A high-frequency restriction fragment length polymorphism was evident in the DNA from 29 unrelated individuals using the enzyme BglII. Analysis of total genomic DNA by digestion with several restriction enzymes, Southern blotting, and hybridization with the human osteopontin cDNA indicated that the gene is a single copy with an approximate length of 5.4-8.2 kb.
AB - A human osteopontin (OP) cDNA was isolated from a library made from primary cultures of human bone cells. The distribution of osteopontin mRNA in human tissues was investigated by Northern analysis and showed that the human message was predominant in cultures of bone cells and in decidua cells isolated at 6-12 weeks of gestation. Immunohistochemical analysis confirmed that OP expression is high in decidua cells as well as in the endometrial glands of a non-pregnant secretory-phase human uterus. Two variants of the OP message were evident on the basis of DNA sequencing and polymerase chain reaction amplification of bone and decidua cell mRNA. The peptides potentially translated by the variant messages differ by the presence (OP1b) or absence (OP1a) of 14 amino acids at residue 58 of the molecule. The deduced human protein sequence shows a conservation between species in the position of the Arg-Gly-Asp (RGD) cell attachment site. Chromosomal mapping of the osteopontin gene (OPN) using human-rodent cell hybrids demonstrated a location on chromosome 4 in the human genome. In situ hybridization of metaphase chromosomes using radiolabeled OP1a as a probe indicated that the gene is located on a region of 4q that is near the centromere. A high-frequency restriction fragment length polymorphism was evident in the DNA from 29 unrelated individuals using the enzyme BglII. Analysis of total genomic DNA by digestion with several restriction enzymes, Southern blotting, and hybridization with the human osteopontin cDNA indicated that the gene is a single copy with an approximate length of 5.4-8.2 kb.
KW - Amino Acid Sequence
KW - Animals
KW - Base Sequence
KW - Cattle
KW - Chromosome Mapping
KW - Chromosomes, Human, Pair 4
KW - Cloning, Molecular
KW - DNA
KW - Decidua
KW - Endometrium
KW - Female
KW - Genes
KW - Humans
KW - Hybrid Cells
KW - Molecular Sequence Data
KW - Osteopontin
KW - Polymerase Chain Reaction
KW - Polymorphism, Restriction Fragment Length
KW - Pregnancy
KW - RNA, Messenger
KW - Rats
KW - Sialoglycoproteins
M3 - Journal article
C2 - 1974876
VL - 7
SP - 491
EP - 502
JO - Genomics
JF - Genomics
SN - 0888-7543
IS - 4
ER -
ID: 34326256