[Expression of human granulysin in murine macrophages and its effects on cell damage and apoptosis].
OBJECTIVE
To construct an eukaryotic expression vector pBudCE4.1/GLS encoding human granulysin(GLS) derived from activated CTLs and observe its effects on cell damage and apoptosis when expressed in macrophages of various murine germ lines.


METHODS
The gene sequence coding GLS carried on pEGFP-C1/GLS was subcloned into pBudCE4.1 plasmid and then transfected into different murine macrophages. The expression of GLS was detected by RT-PCR and immunocytochemical staining. The impairment and apoptosis of the host cells was assesed by measuring LDH in culture media and fluorescent staining of nuclear DNA, respectively.


RESULTS
The pBudCE4.1/GLS recombinant containing accurate ORF of GLS was obtained and successfully expressed in the target cells at 48 h after transfection. No obvious effects on cell damage and apoptosis was detected during this course.


CONCLUSIONS
The gene coding human GLS can be expressed in murine macrophages. The expressed product has no obvious effects on cell damage and apoptosis.
Yi
Zheng-jun
ZJ
Department of Microbiology and Immunology, Chongqing University of Medical Sciences, Chongqing 400016, China. yizhengjun@sohu.com
Zhu
Dao-yin
DY
Li
Jun-ming
JM
Yang
Jian
J
He
Yong-lin
YL
Zeng
Wei
W
Li
Na
N
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics* / Q000378:metabolism*; D017209:Apoptosis; D002460:Cell Line; D006801:Humans; D007150:Immunohistochemistry; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000945:Antigens, Differentiation, T-Lymphocyte; C075005:GNLY protein, human