Using Lab On-line to
Clone and Identify the Esophageal Cancer Related Gene 4
BI Mei-Xia， HAN Wei-Dong1, LU Shi-Xin*
( Cancer Institute(Hospital), Chinese Academy of Medical Sciences, Peiking
Union Medical College, Beijing 100021, China; 1Institute
of Basic Medical Science, General Hospital of the Chinese Peoples Liberation
Army, Beijing 100853, China )
Abstract Using
Internet as platform, databases as materials and software as tools to assemble
a lab on-line is revolutionizing in bioscience research. The major works of lab
on-line are cloning, identification, localization of genes, and the structural
and functional analysis of proteins. In this report, the esophageal cancer
related gene 4(ECRG-4)(accession number： AF325503) was successfully
isolated. The 97 bp ECRG-4 EST was initiallyusedto fish the human EST
databases. Five pieces of ESTs showed strong homology to it, and they were
assembled to one 772 bp cDNA sequence by DNASTAR software. Then the 447 bp full
open reading frame of ECRG-4 was determined by ORF FINDERto encode 148
amino acids. Sequence of ECRG-4 did not reveal remarkable similarity
tothe known sequences in a homology analysis with the public database of
GenBank,showing that it is a new gene. Homology analysis of protein coding by ECRG-4
showed a 31% homology with mouse IgG V region. ECRG-4 gene is
expressed in normal esophagus, bladder and brain tissues, but its expression
was significantly down regulated in prostate tumors and tumor cell lines. ECRG-4
gene was located in 2q14.1–4.3 by HTGS and STS, and was conformed by radiation
hybrid (RH) method. We propose that this purely lab on line cloning approach
can be used by modestly sized laboratories to rapidly and accurately
characterize human genes without wasting too much money and time.
Key words lab on-line； gene cloning； ECRG-4
*Corresponding author： Tel, 86-10-67712368； Fax， 86-10-67713359； e-mail, [email protected]