Molecular cloning of DNA complementary to bovine adrenal P450scc mRNA

Karla J. Matteson; Bon chu Chung; Walter L. Miller

doi:10.1016/0006-291X(84)91443-8

Molecular cloning of DNA complementary to bovine adrenal P450_scc mRNA

Karla J. Matteson, Bon chu Chung, Walter L. Miller

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

11 Citations (Scopus)

Abstract

P450_scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450_scc indicates P450_scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the PstI site of pBR322 by dC·dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450_scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert.

Original language	English
Pages (from-to)	264-270
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	120
Issue number	1
DOIs	https://doi.org/10.1016/0006-291X(84)91443-8
Publication status	Published - Apr 16 1984
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1016/0006-291X(84)91443-8

Cite this

@article{f963ac767b504069b94e8f936b6aca42,

title = "Molecular cloning of DNA complementary to bovine adrenal P450scc mRNA",

abstract = "P450scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450scc indicates P450scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the PstI site of pBR322 by dC·dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert.",

author = "Matteson, {Karla J.} and Chung, {Bon chu} and Miller, {Walter L.}",

note = "Funding Information: We thank Drs. Michael R. Waterman and Evan R. Siqson of the University of Texas Health Science Center in Dallas, who provided us with the rabbit anti-bovine P450 antibody without which we could not have done this work (supported by N#&ant AM 28350 to M.R.W.). We also thank John Fiddes for advice about the transformation procedure and Harald Biesmnann for his gift of --E. coli MM 294. Supported by NIH grant HD 16047 to W.L.M.",

year = "1984",

month = apr,

day = "16",

doi = "10.1016/0006-291X(84)91443-8",

language = "English",

volume = "120",

pages = "264--270",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Molecular cloning of DNA complementary to bovine adrenal P450scc mRNA

AU - Matteson, Karla J.

AU - Chung, Bon chu

AU - Miller, Walter L.

N1 - Funding Information: We thank Drs. Michael R. Waterman and Evan R. Siqson of the University of Texas Health Science Center in Dallas, who provided us with the rabbit anti-bovine P450 antibody without which we could not have done this work (supported by N#&ant AM 28350 to M.R.W.). We also thank John Fiddes for advice about the transformation procedure and Harald Biesmnann for his gift of --E. coli MM 294. Supported by NIH grant HD 16047 to W.L.M.

PY - 1984/4/16

Y1 - 1984/4/16

N2 - P450scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450scc indicates P450scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the PstI site of pBR322 by dC·dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert.

AB - P450scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450scc indicates P450scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the PstI site of pBR322 by dC·dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert.

UR - http://www.scopus.com/inward/record.url?scp=0021765230&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021765230&partnerID=8YFLogxK

U2 - 10.1016/0006-291X(84)91443-8

DO - 10.1016/0006-291X(84)91443-8

M3 - Article

C2 - 6546877

AN - SCOPUS:0021765230

SN - 0006-291X

VL - 120

SP - 264

EP - 270

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 1

ER -

Molecular cloning of DNA complementary to bovine adrenal P450_scc mRNA

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this