Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain

Mon Li Chu; Jeanne C. Myers; Michael P. Bernard; Juy Fang Ding; Francesco Ramirez

doi:10.1093/nar/10.19.5925

Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain

Mon Li Chu
, Jeanne C. Myers
, Michael P. Bernard
, Juy Fang Ding
, Francesco Ramirez

Research output: Contribution to journal › Article › peer-review

423 Scopus citations

Abstract

We report the cloning of five overlapping cDNAs bearing sequences specific for the human proα1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endo-nuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the α chain to part of the 3′ end untranslated region of the proα1(I) mRNA for a total of 3400 nucleotides.

Original language	English
Pages (from-to)	5925-5934
Number of pages	10
Journal	Nucleic Acids Research
Volume	10
Issue number	19
DOIs	https://doi.org/10.1093/nar/10.19.5925
State	Published - 11 Oct 1982
Externally published	Yes

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@article{1e3a65a46e204138899615cd5ee47f9e,

title = "Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain",

abstract = "We report the cloning of five overlapping cDNAs bearing sequences specific for the human proα1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endo-nuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the α chain to part of the 3′ end untranslated region of the proα1(I) mRNA for a total of 3400 nucleotides.",

author = "Chu, \{Mon Li\} and Myers, \{Jeanne C.\} and Bernard, \{Michael P.\} and Ding, \{Juy Fang\} and Francesco Ramirez",

note = "Funding Information: siastic support and for many useful discussions and helpful suggestions. This work was supported by grants from NIH (AM-16516) and the Lalor Foundation.",

year = "1982",

month = oct,

day = "11",

doi = "10.1093/nar/10.19.5925",

language = "English",

volume = "10",

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journal = "Nucleic Acids Research",

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TY - JOUR

T1 - Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain

AU - Chu, Mon Li

AU - Myers, Jeanne C.

AU - Bernard, Michael P.

AU - Ding, Juy Fang

AU - Ramirez, Francesco

N1 - Funding Information: siastic support and for many useful discussions and helpful suggestions. This work was supported by grants from NIH (AM-16516) and the Lalor Foundation.

PY - 1982/10/11

Y1 - 1982/10/11

N2 - We report the cloning of five overlapping cDNAs bearing sequences specific for the human proα1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endo-nuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the α chain to part of the 3′ end untranslated region of the proα1(I) mRNA for a total of 3400 nucleotides.

AB - We report the cloning of five overlapping cDNAs bearing sequences specific for the human proα1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endo-nuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the α chain to part of the 3′ end untranslated region of the proα1(I) mRNA for a total of 3400 nucleotides.

UR - https://www.scopus.com/pages/publications/0020480644

U2 - 10.1093/nar/10.19.5925

DO - 10.1093/nar/10.19.5925

M3 - Article

C2 - 6183642

AN - SCOPUS:0020480644

SN - 0305-1048

VL - 10

SP - 5925

EP - 5934

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 19

ER -

Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain

Abstract

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