Rival S, Attal J, Delville-Giraud C, Yerle M, Laffont P, Rogel-Gaillard C, Houdebine L
Laboratoire de Biologie du Développement et Biotechnologies, Institut National de Recherche Agronomique, 78352 Jouy en Josas cedex, Cedex, France.
Gene. 2001 Apr 4;267(1):37-47. doi: 10.1016/s0378-1119(01)00388-2.
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.
乳清酸性蛋白（WAP）是啮齿动物、兔子和骆驼的主要乳清蛋白。最近，在猪的乳汁中也发现了这种蛋白（Simpson等人，1998年。《分子内分泌学杂志》20卷，27 - 35页）。本文报道了猪WAP cDNA的克隆以及包含整个猪WAP基因的细菌人工染色体（BAC）构建体。将猪WAP基因的编码序列与已发表的啮齿动物或兔形目动物的WAP序列进行比较，结果表明只有外显子序列部分保守。猪WAP基因定位于18号染色体的亚端粒区域。对泌乳动物乳腺中猪WAP基因表达的估计显示其表达水平很高。为了比较包含猪WAP基因下游70 kb和上游50 kb的大片段基因组或较小片段（下游1 kb和上游2.4 kb）中猪WAP基因的表达水平，将这两个基因组片段转染到HC11细胞系中。与整合拷贝数相比，BAC构建体的表达比质粒高15倍。本报告表明，HC11细胞系是鉴定乳蛋白基因调控序列的有用工具。
