[Preparation of anti-Red antisera and its subcellular localization].
OBJECTIVE
To prepare rabbit anti-Red antisera.


METHODS
The bet, exo and gam genes of lambda phage were amplified by PCR from genomic DNA and cloned into the expression vector pDH2, respectively. Red proteins were induced to express at 42 degrees C. The expressed proteins were analyzed by PAGE and thin-layer scanning. The antisera were prepared by immunizing rabbits with the three Red proteins, respectively. The titers and specificities of the antisera were detected by Western blot.


RESULTS
Beta, Exo and Gam proteins accounted for about 40.3%, 49.2% and 73.4% of total bacterial protein, respectively. The titers of the antisera were about 1:2,000. Western blot analysis indicated that the three antisera all had good specificities to the corresponding proteins.


CONCLUSIONS
Specific anti-Red antisera are prepared successfully.
Zhang
Xiao-nan
XN
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Chen
Su-min
SM
Luo
Er-ping
EP
Huang
Yong
Y
Chai
Yu-bo
YB
Li
Yun-feng
YF
Chen
Nan-chun
NC
D000818:Animals; D010582:Bacteriophage lambda / Q000276:immunology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering; D007106:Immune Sera / Q000032:analysis* / Q000276:immunology*; D042541:Intracellular Space / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D007106:Immune Sera; D014764:Viral Proteins