An integrated technique for identification of differential genes expressed in patients with cancer.
To develop a method for identification of differential gene expression between different cell populations, several convenient techniques of molecular biology, including subtractive hybridization, suppression PCR, T/A cloning and sequencing, were used to identify genes expressed differentially in CD45+ and CD45- cells isolated from U266 cell line of multiple myeloma. Our results showed that the levels of abundant genes scale down 20 times through subtractive hybridization. Plasmid DNA from CD45- cell clones was hybridized with forward or backward cDNA probes synthesized from CD45- and CD45- cells, respectively. A few of differentially expressed genes reconfirmed by RT-PCR were identified from 500 expressed clones of CD45+ cells. It is concluded that a strategy for gene expression identification developed from conventional molecular biological methods can be used in different laboratories.
Li
Shouxin
S
Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Liu
Shangqin
S
Lei
Xiaomei
X
D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D017493:Leukocyte Common Antigens / Q000235:genetics*; D009101:Multiple Myeloma / Q000235:genetics / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004247:DNA; D017493:Leukocyte Common Antigens