Identification of a novel regeneration-related gene H3 and its protein from the differential expression cDNA library of spinal cord injury in neonatal rats.
- Author:
Tong-wei CHU
1
;
Yu-gang LIU
;
Hai-han MA
;
Wei-hong LIAO
;
Ya-min WU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Blotting, Northern; DNA, Complementary; genetics; Gene Library; Molecular Sequence Data; Nerve Regeneration; genetics; Rats; Reverse Transcriptase Polymerase Chain Reaction; Spinal Cord Injuries; genetics
- From: Chinese Journal of Traumatology 2007;10(4):195-199
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo clone and identify one novel regeneration related gene H(3) (CA854305) from the differential expression genes library we had set up before.
METHODSUse the method of Northern blot to detect the different expressions of the novel gene under different situations, employ the technique of in silico cloning to scan the span of the novel gene, and analyze their sequences. Also we used reverse transcription PCR to validate the largest open reading frame.
RESULTSNorthern blotting results of H(3) (CA854305) showed that the transplanted group had more efficient and extensive expression than untreated and uninjured groups 5 days after spinal cord injury, while the untreated group had more extensive expression than uninjured group. It implied that H(3) might have some relationship with nerve regeneration after spinal cord injury. From the results of in silico cloning we got a longest contig of 1635 bp and an largest open reading frame of 542 bp from 49 to 591 bp correspondent with the Cozak rules. Reverse transcription PCR validated the largest open reading frame sequence primarily.
CONCLUSIONSWe got the sequence of novel gene H(3) which might be one of the regenerationjrelated genes. Key words:Gene library; Genes; Nerve regeneration; Spinal cord injuries.