Cloning of cDNA to rat mammary-gland fatty acid synthase mRNA. Evidence for the expression of two mRNA species during lactation.
A cDNA library was constructed in the expression vector lambda gt11, by synthesizing cDNA from size-selected poly(A) RNA from lactating rat mammary gland, using random hexanucleotide primers. Using this library we identified two recombinants which, on addition of a lac z inducer, produced proteins recognized by affinity-purified anti-fatty-acid synthase antibody, and which, therefore, contained fatty acid synthase coding sequences. The inserts were subcloned, were shown to be between 500 and 600 base pairs in size, and to cross-hybridize. The cloned DNA was then used in Northern hybridizations with mRNA isolated at various stages throughout lactation. Two mRNA species were identified of approx. 9.7 and 10.4 kilobases, which increased and decreased in parallel during lactation, reaching a peak at 12-13 days. Both mRNA species disappeared rapidly if the pups were removed prematurely. This study provides evidence that, during hormonal induction in lactation, regulation of the level of fatty acid synthase protein can be accounted for by variation in the level of mRNA.
Braddock
M
M
Department of Biochemistry, Dundee University, U.K.
Hardie
D G
DG
D000818:Animals; D000906:Antibodies; D003001:Cloning, Molecular; D004270:DNA, Circular / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D064429:Fatty Acid Synthases / Q000235:genetics* / Q000276:immunology; D005260:Female; D007774:Lactation / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D004270:DNA, Circular; D012333:RNA, Messenger; D064429:Fatty Acid Synthases