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cDNA cloning of new protein tyrosine phosphatases in the human colon
1Department of Internal Medicine (Section 1), Sapporo Medical College, Japan.
Abstract:
Oligonucleotide probes corresponding to three invariant amino acid sequence motifs within the protein tyrosine phosphatase (PTPase) catalytic domain were synthesized and employed in the polymerase chain reaction (PCR). PCR products amplified from human colonic tissues were analyzed. Among 41 cDNA clones sequenced, two putative novel PTPases designated as CL-6 and CL-2B were isolated. cDNA cloning of CL-6 was performed, and three overlapped cDNA clones encoding part of the coding region, which was 2.1 kb long, were obtained. Expression of mRNAs of these clones was then investigated by Northern blot analysis or reverse transcriptase-PCR, suggesting that CL-6 mRNA may be expressed in a cell-type-specific manner, whereas the expression of CL-2B mRNA could be ubiquitous.
Insights
Researchers identified two novel protein tyrosine phosphatases (PTPases), CL-6 and CL-2B, in human colon tissue using polymerase chain reaction (PCR). CL-6 shows cell-specific expression, while CL-2B appears ubiquitous.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Protein tyrosine phosphatases (PTPases) play critical roles in cellular signaling.
- Identifying novel PTPases is essential for understanding diverse biological processes.
Purpose of the Study:
- To isolate and characterize novel PTPases from human colonic tissues.
- To investigate the expression patterns of newly identified PTPases.
Main Methods:
- Oligonucleotide probes targeting conserved PTPase motifs were designed.
- Polymerase chain reaction (PCR) was used to amplify target sequences from human colonic cDNA.
- cDNA cloning, sequencing, Northern blot analysis, and reverse transcriptase-PCR were employed for characterization and expression analysis.
Main Results:
- Two novel PTPases, designated CL-6 and CL-2B, were successfully isolated from 41 sequenced cDNA clones.
- Full-length cDNA cloning of CL-6 yielded a 2.1 kb coding region.
- CL-6 mRNA exhibited cell-type-specific expression, whereas CL-2B mRNA demonstrated ubiquitous expression.
Conclusions:
- The study successfully identified two novel PTPases, CL-6 and CL-2B, contributing to the PTPase family.
- Differential expression patterns suggest distinct functional roles for CL-6 and CL-2B in human tissues.