Analysis of the flanking regions of the human alpha-lactalbumin gene responsible for position-effect independent expression.
Transgenic rats with the 130 kb bacterial artificial chromosome construct bLA, including the alpha-lactalbumin gene, had position-independent and copy number-dependent expression, which confirmed previous experiments using the 210 kb yeast artificial construct, yLALBA. To identify elements that confer a position effect, we compared the yLALBA and bLA sequences. yLALBA was chimeric. A common 32 kb region was identified and the total nucleotide sequence was determined. We previously analyzed transgenic rats using polymerase chain reaction to compare the integrity and expression of the transgenes. The -6 to +9 kb region is considered to be necessary for position-independent expression. Transgenic rats lacking the -3.4 to -0.85 kb region had a severe position effect. This 2.5 kb region contains two DNaseI hypersensitive sites at -1.0 and -2.8 kb. The 2.5 kb region is proposed to be a locus control region of the human alpha-lactalbumin gene.
Fujiwara
Yoshihiro
Y
Division of Immunology, National Institute of Animal Health, Kannondai 3-1-1, Tsukuba, Ibaragi 305-0856, Japan.
Takahashi
Ri-ichi
R
Hirabayashi
Masumi
M
Ueda
Masatsugu
M
Muramatsu
Tatsuo
T
Yamanaka
Harumichi
H
Sekikawa
Kenji
K
D024506:5' Flanking Region / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites / Q000235:genetics; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D018628:Gene Dosage; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D007768:Lactalbumin / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D017422:Sequence Analysis, DNA
D004247:DNA; D007768:Lactalbumin; D003850:Deoxyribonuclease I