Cloning, transcription and chromosomal localization of the porcine whey acidic protein gene and its expression in HC11 cell line.
The whey acidic protein (WAP) is the major whey protein of rodent, rabbit and camel. Recently, it was identified in the milk of swine (Simpson et al., 1998. J. Mol. Endocrinol. 20, 27-35). In this paper, the cloning of the pig WAP cDNA and of bacterial artificial chromosome (BAC) construct containing the entire porcine WAP gene is reported. The comparison of the coding sequence of the pig WAP gene to rodent or lagomorph WAP sequence already published demonstrated that only exon sequences are partially conserved. The porcine WAP gene was localized on the subtelomeric region of the chromosome 18. The estimation of the expression of the swine WAP gene in the mammary gland from lactating animals revealed a high level of expression. In order to compare the expression level of the porcine WAP gene from the large genomic fragment which contained 70 kb downstream and 50 kb upstream the pig WAP gene or the smaller one (1 kb downstream and 2.4 kb upstream), these two genomic fragments were transfected in HC11 cell line. The BAC construct was expressed 15 times higher than the plasmid when reported to the integrated copy number. This report suggests that the HC11 cell line is a useful tool to identify the regulatory sequences of milk protein genes.
Rival
S
S
Laboratoire de Biologie du Développement et Biotechnologies, Institut National de Recherche Agronomique, 78352 Jouy en Josas cedex, Cedex, France.
Attal
J
J
Delville-Giraud
C
C
Yerle
M
M
Laffont
P
P
Rogel-Gaillard
C
C
Houdebine
L
L
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002871:Chromosome Banding; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D005796:Genes / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D007774:Lactation; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008894:Milk Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D012313:RNA / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014158:Transcription, Genetic
D008894:Milk Proteins; C024471:whey acidic proteins; D012313:RNA; D004247:DNA