An integrated technique for identification of differential genes expressed in patients with cancer.
- Author:
Shouxin LI
1
;
Shangqin LIU
;
Xiaomei LEI
Author Information
1. Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
DNA;
genetics;
Gene Expression Profiling;
Gene Expression Regulation, Neoplastic;
Humans;
Leukocyte Common Antigens;
genetics;
Multiple Myeloma;
genetics;
pathology;
Reverse Transcriptase Polymerase Chain Reaction
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2004;24(6):578-580
- CountryChina
- Language:English
-
Abstract:
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.
