[A new strategy of gene therapy for hyperphenylalaninemia rats].
OBJECTIVE
To construct a recombinant vector which expresses active phenylalanine-amonia-lyase (PAL) in Lactococcus lactis (L. L.), and to convert phe into cinnamic acid in small intestine by the engineering L. L. to decrease the phe level in the peripheral blood, and to cure hyperphenylalaninemia rats.


METHODS
PAL cDNA from Petroselinum crispum was subcloned into expression vector pMG36e and transformed L. L. The pMG36ePAL/L.L. was screened and characterized by using PCR and HPLC, and prepared as enteric-coated microcapsules and oral liquid type preparation that were given orally to hyperphenylalaninemia-rats.


RESULTS
Engineering L. L. expressing PAL activity was obtained. The phe levels plasma of in the rats receiving preparations made from the engineering L. L. were significantly reduced compared with non-treated hyperphenylalaninemia rats. And the effects of different preparations were different from each other.


CONCLUSIONS
The engineering L. L. expressing PAL activity can reduce the blood phe level of the hyperphenylalaninemia rats. This may be a potential way for PKU gene therapeutics.
Jia
X
X
Beijing Red-Cross Chaoyang Hospital, Capital University of Medical Sciences Beijing, 100020 China.
Liu
J
J
Xiang
H
H
D000284:Administration, Oral; D000818:Animals; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D013294:Lactococcus lactis / Q000235:genetics*; D008297:Male; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis / Q000235:genetics*; D010661:Phenylketonurias / Q000628:therapy*; D051381:Rats; D016896:Treatment Outcome
D010650:Phenylalanine Ammonia-Lyase