NDLI: Transgenic Mice Can Express Mutant Human Coagulation Factor IX with Higher Level of Clotting Activity

Content Provider	Springer Nature Link
Author	Yan, Jing Bin Wang, Shu Huang, Wen Ying Xiao, Yan Ping Ren, Zhao Rui Huang, Shu Zheng Zeng, Yi Tao
Copyright Year	2006
Abstract	To improve the available values of transgenic animals, we produced a mutant human coagulation factor IX minigene (including cDNA and intron I) with arginine at 338 changed to alanine (R338A-hFIX) by using a direct mutation technique. The R338A-hFIX minigene was then cloned into a plasmid carrying the goat β-casein promoter to get a mammary gland-specific expression vector. The clotting activity in the supernatant of the transfected HC-11 cells increased to approximately three times more than that of wild-type hFIX. Nine transgenic mice (three females and six males) were produced, and the copy number of the foreign gene was very different, ranging from 1 to 43 in different lines. ELISA, Western blot, and clotting assay experiments showed that the transgenic mice could express R338A-hFIX, showing higher average levels of clotting activity than wild-type hFIX in the milk (103.76% vs. 49.95%). The highest concentration and clotting activity of hFIX reached 26 μg/mL and 1287% in one founder (F0-7), which was over 10 times higher than that in human plasma. Furthermore, RT-PCR, APTT assay, and histological analysis indicated that hFIX was expressed specifically in the mammary gland without affecting the intrinsic coagulation pathway and physiologic performance of the local tissue.
Starting Page	347
Ending Page	358
Page Count	12
File Format	PDF
ISSN	00062928
Journal	Biochemical Genetics
Volume Number	44
Issue Number	7-8
e-ISSN	15734927
Language	English
Publisher	Kluwer Academic Publishers-Plenum Publishers
Publisher Date	2006-09-21
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Medical Microbiology Zoology Biochemistry Human Genetics
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Biochemistry Molecular Biology Ecology, Evolution, Behavior and Systematics

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