Large-scale purification and acute toxicity of hygromycin B phosphotransferase.
OBJECTIVE
To provide the acute toxicity data of hygromycin B phosphotransferase (HPT) using recombinant protein purified from E. coli.


METHODS
Recombinant HPT protein was expressed and purified from E. coli. To exclude the potential adverse effect of bacteria protein in recombinant HPT protein, bacterial control plasmid was constructed, and bacteria control protein was extracted and prepared as recombinant HPT protein. One hundred mice, randomly assigned to 5 groups, were administrated 10 g/kg, 5 g/kg, or 1 g/kg body weight of HPT or 5 g/kg body weight of bacterial control protein or phosphate-buffered saline (PBS) respectively by oral gavage.


RESULTS
All animals survived with no significant change in body weight gain throughout the study. Macroscopic necropsy examination on day 15 revealed no gross pathological lesions in any of the animals. The maximum tolerated dose (MTD) of HPT was 10 g/kg body weight in mice and could be regarded as nontoxic.


CONCLUSIONS
HPT protein does not have any safety problems to human health.
Zhuo
Qin
Q
Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Piao
Jian-Hua
JH
Tian
Yuan
Y
Xu
Jie
J
Yang
Xiao-Guang
XG
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000633:toxicity*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000302:isolation & purification / Q000633:toxicity*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000633:toxicity*; D023382:Toxicity Tests, Acute
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase