AIM: To isolate and analyze a novel gene over-expressed during liver regeneration. METHODS: Total RNA of regenerating liver was extracted from liver tissue after 0-4-36-36-36 hr short interval successive partial hepatectomy (SISPH). Reverse transcription-polymerase chain reaction was used to synthesize double strand cDNA, after the tissue was digested by proteinase K and Sfi A/B. The double-strand cDNA was ligated to lambdaTriplEx2. lambdaphage packaging reaction was performed and E. coli XL1-Blue was infected for titering and amplifying. One expressed sequence tag was probed by Dig and phage in situ hybridization was carried out to isolate positive clones. Positive recombinant lambdaTriplEx2 was converted to the corresponding pTriplEx2, and bioinformatics was used to analyze full-length cDNA. RESULTS: We isolated a novel full-length cDNA during liver regeneration following SISPH. CONCLUSION: We have succeeded in cloning a novel gene, based on bioinformatics. We postulate that this gene may function in complicated network in liver regeneration. On the one hand, it may exert initiation of liver regeneration via regulating nitric oxide synthesis. On the other hand, it may protect damaged residue lobus following SISPH.
AIM: To isolate and analyze a novel gene over-expressed during liver regeneration. METHODS: Total RNA of regenerating liver was extracted from liver tissue after 0-4-36-36-36 hr short interval successive partial hepatectomy (SISPH). Reverse transcription-polymerase chain reaction was used to synthesize double strand cDNA, after the tissue was digested by proteinase K and Sfi A/B. The double-strand cDNA was ligated to lambdaTriplEx2. lambdaphage packaging reaction was performed and E. coli XL1-Blue was infected for titering and amplifying. One expressed sequence tag was probed by Dig and phage in situ hybridization was carried out to isolate positive clones. Positive recombinant lambdaTriplEx2 was converted to the corresponding pTriplEx2, and bioinformatics was used to analyze full-length cDNA. RESULTS: We isolated a novel full-length cDNA during liver regeneration following SISPH. CONCLUSION: We have succeeded in cloning a novel gene, based on bioinformatics. We postulate that this gene may function in complicated network in liver regeneration. On the one hand, it may exert initiation of liver regeneration via regulating nitric oxide synthesis. On the other hand, it may protect damaged residue lobus following SISPH.
Authors: G D Block; J Locker; W C Bowen; B E Petersen; S Katyal; S C Strom; T Riley; T A Howard; G K Michalopoulos Journal: J Cell Biol Date: 1996-03 Impact factor: 10.539