LIVER-SPECIFIC EXPRESSION OF A PHOSPHOENOLPYRUVATE CARBOXYKINASE-NEO GENE IN GENETICALLY MODIFIED CHICKENS

被引:7
作者
COOK, RF
COOK, SJ
SAVON, S
MCGRANE, M
HARTITZ, M
HANSON, RW
HODGSON, CP
机构
[1] OHIO STATE UNIV, OHIO AGR RES & DEV CTR, MOLEC BIOL LAB, WOOSTER, OH 44691 USA
[2] OHIO STATE UNIV, OHIO AGR RES & DEV CTR, DEV BIOL LAB, WOOSTER, OH 44691 USA
[3] CASE WESTERN RESERVE UNIV, SCH MED, DEPT BIOCHEM, CLEVELAND, OH 44106 USA
关键词
GENETIC MANIPULATION; RETROVIRAL VECTORS; PHOSPHOENOLPYRUVATE CARBOXYKINASE GENE PROMOTER; GENE THERAPY;
D O I
10.3382/ps.0720554
中图分类号
S8 [畜牧、 动物医学、狩猎、蚕、蜂];
学科分类号
0905 ;
摘要
In order to investigate the potential of the avian liver for the expression of recombinant proteins in vivo, replication-competent retroviral vectors were used to introduce a recombinant rat phosphoenolpyruvate carboxykinase promoter-driven neomycin resistance gene (PEPCKneo) into early Line 11 Leghorn embryos. After hatching, these birds possessed apparently intact PEPCKneo sequences in most tissues examined, however, the neo protein was expressed preferentially in the liver (up to .45% of total cellular protein). Therefore, the tissue specificity of the PEPCK promoter from the rat was retained in the chicken, although hormone responsiveness was not observed. Retroviral vectors used to transmit the genes were more stable during passage in either fibroblast cells or in the animal if the inserted genes were oriented in the same (sense) direction as the viral genome. After Geneticin drug selection in cultured cells, PEPCKneo mRNA was the predominant recombinant species observed on Northern blots, whereas embryos expressed mostly the RNA species originating in the retroviral long terminal repeats. The results demonstrate the potential usefulness of liver-specific gene expression in chickens, as well as the transcriptional effects observed when a foreign promoter is introduced into the replication-competent vector.
引用
收藏
页码:554 / 567
页数:14
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