[The establishment of conditional MT transgenic mice model].
OBJECTIVE
To construct the doxycycline-inducible MT transgenic mice model, and provide a basis for the study of hemangioma as well as MT molecular function in vivo.


METHODS
Tetracycline-controlled expression systems were employed to this study. A conditional transgenic vector combining the two transcriptional units on a single plasmid was constructed, and the MT gene was subcloned into this vector. To minimize any potential interference, the two elements were spaced with a 1.2 kb cHS4 insulator. To shield the transgene from the affection of chromosomal position effect and improve its expression efficiency, another cHS4 insulator was inserted into the upstream of transgene cassette. After transient transfection of cells in vitro, and analyzing the relative quantification of MT transcripts (target) in mRNA samples by semi-quantitative RT-PCR method, the pronuclear microinjection technique was used to introduce the purified transgene into the chromosomes of fertilized mice eggs, in order to obtain transgenic positive animals. The MT expression in positive mouse was induced through adding deoxycycline in drinking water. Phenotype analysis was done by pathology, and MT expression was confirmed by RT-PCR.


RESULTS
The conditional transgenic vector was constructed successfully, and the expression of MT in vitro was regulated by doxycycline. Five transgenic positive mice were obtained through pronuclear microinjection. After MT induction, one transgenic mice developed hemangiomas, and the expression of MT was confirmed by RT-PCR method. The others were active and in breeding.


CONCLUSIONS
Conditional MT transgenic animal model was constructed successfully, and may provide platform for the experimental research of hemangioma as well as the MT molecular function in vivo.
Wang
Yan-an
YA
Department of Oral & Maxillofacial Surgery, Ninth People's Hospital, School of Stomatology, Shanghai Jiaotong University, Shanghai, 200011 PR China.
Zheng
Jia-wei
JW
Fei
Zhao-liang
ZL
Jiang
Xin-quan
XQ
Li
Jin-zhong
JZ
Xu
Qin
Q
Wang
Zhu-gang
ZG
Fei
Jian
J
Zhang
Zhi-yuan
ZY
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D019556:COS Cells; D002522:Chlorocebus aethiops; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013752:Tetracycline / Q000494:pharmacology*; D014162:Transfection / Q000379:methods
D000952:Antigens, Polyomavirus Transforming; D013752:Tetracycline