[Discovery of rat Islet-1 gene variant and its expression in NSC].
OBJECTIVE
To construct rat Islet-1 gene recombinant retroviral expression vector and to transduce Islet-1 gene into neural stem cell (NSC) by the vector.


METHODS
The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method, the amplified gene fragment was subcloned into the retroviral vector plEGFP-C1. Islet-1 gene was transduced into NSC by PA317 packaging cells, then the expression of Islet-1 in NSC was observed.


RESULTS
The recombinant Islet-1 retroviral vector was constructed successfully, as shown by PCR, restriction enzymes digestion and fluorescence detection, and the positive expression of Islet-1 was found in NSC by immunocytochemical staining. The variant of Islet-1 gene was also discovered in this study.


CONCLUSIONS
The rat Islet-1 recombinant retroviral vector has been successfully prepared, which can be used to study the function of Islet-1 gene in NSC differentiation into cholinergic neurons.
Liu
Jia-mei
JM
Department of Histology and Embryology, School of Basic Medical Science, Jilin University, Changchun 130021, China. liujiamei100@yahoo.com.cn
Chen
Dong
D
Meng
Xiao-ting
XT
D000818:Animals; D002460:Cell Line; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism*; D060850:LIM-Homeodomain Proteins; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors; D014162:Transfection
D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; D014157:Transcription Factors; C063843:insulin gene enhancer binding protein Isl-1