[Screening of aberrant genes of immunocytes in severely scalded rats by inhibition of subtractive hybridization].
OBJECTIVE
To explore the relationship between the postburn changes of gene expression of immunocytes and postburn immunological functional disturbance in severely scalded rats.


METHODS
Immunocytes of female F344 rats with severe scalding were harvested and the mRNA was extracted for the inhibition of the expression of aberrant gene selected by subtractive hybridization in contrast to that before scalding. The aberrant genes were cloned and sequenced and compared to those in Genebank for homology.


RESULTS
The harvested immunocytes were identified by LM to be monocytes and lymphocytes. Multiple strips located at 200 approximately 400 bp were obtained by inhibiting subtractive hybridization and were cloned into PGEM-T easy plasmid vector. Thus the genebank of aberrant expression was constructed. Twenty clones were sequenced randomly and partial obtained known genes were in accord with postburn systemic changes. Several novel genes were accepted and registered by Genebank.


CONCLUSIONS
There exhibited change in the gene expression of peripheral immunocytes at early postburn stage in rats inflicted by severe burn.
Shi
S
S
Department of Burns, Changhai Hospital, Second Military Medical University, Shanghai 200433, P. R. China.
Xia
Z
Z
D000818:Animals; D002056:Burns / Q000235:genetics / Q000473:pathology*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation; D007107:Immune System / Q000166:cytology / Q000378:metabolism*; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D012333:RNA, Messenger