TY - JOUR
T1 - Nebulin cDNAs detect a 25-kilobase transcript in skeletal muscle and localize to human chromosome 2
AU - Stedman, Hansell
AU - Browning, Karen
AU - Oliver, Noelynn
AU - Oronzi-Scott, Marion
AU - Fischbeck, Kenneth
AU - Sarkar, Satyapriya
AU - Sylvester, James
AU - Schmickel, Roy
AU - Wang, Kuan
N1 - Funding Information:
We thank J. Shipsey, G. Gutierrex, Richard Petersen, and D. Sosnoski for excellent technical assistance, R. Nussbaum for the gift of human-hamster hybrid DNA preparations, A. Burghes and R. Worton for provision of cDNA JK1.4, and W. Meredith for dedicated preparation of the manuscript and figures. This work was supported by grants from the Muscular Dystrophy Association (J.E.S.) and National Institutes of Health Grant KD-20270 (K.W.). H.S. is the recipient of the Robert G. Sampson Postdoctoral Fellowship from the MDA. K.W. is the recipient of an American Heart Association Established Investigatorship.
PY - 1988/1
Y1 - 1988/1
N2 - By virtue of the protein's size, myofibrillar localization, and proposed functional role, the gene encoding the giant sarcomere matrix protein nebulin represents a possible site for myopathic mutations. Using polyclonal anti-nebulin antisera to screen a cDNA expression library, we have isolated and characterized two separate human fetal muscle cDNA clones. By recovering fusion polypeptide-bound portions of our polyclonal antiserum and reutilizing them to probe Western blots, we further demonstrate that the expressed cDNAs encode polypeptide epitopes unique to the protein nebulin. Both cDNAs detect a 25-kb skeletal muscle RNA transcript and localize to human chromosome 2. The identification of nebulin cDNA clones enables the complete analysis of this enormous mRNA by transcript walking through muscle cDNA libraries. Here we report a restriction map of the 3′ end of the human nebulin transcript, with reference to the genomic fragments identified by the cDNA.
AB - By virtue of the protein's size, myofibrillar localization, and proposed functional role, the gene encoding the giant sarcomere matrix protein nebulin represents a possible site for myopathic mutations. Using polyclonal anti-nebulin antisera to screen a cDNA expression library, we have isolated and characterized two separate human fetal muscle cDNA clones. By recovering fusion polypeptide-bound portions of our polyclonal antiserum and reutilizing them to probe Western blots, we further demonstrate that the expressed cDNAs encode polypeptide epitopes unique to the protein nebulin. Both cDNAs detect a 25-kb skeletal muscle RNA transcript and localize to human chromosome 2. The identification of nebulin cDNA clones enables the complete analysis of this enormous mRNA by transcript walking through muscle cDNA libraries. Here we report a restriction map of the 3′ end of the human nebulin transcript, with reference to the genomic fragments identified by the cDNA.
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U2 - 10.1016/0888-7543(88)90102-4
DO - 10.1016/0888-7543(88)90102-4
M3 - Article
C2 - 2838409
AN - SCOPUS:0023758207
SN - 0888-7543
VL - 2
SP - 1
EP - 7
JO - Genomics
JF - Genomics
IS - 1
ER -