Identification of a novel regeneration-related gene H3 and its protein from the differential expression cDNA library of spinal cord injury in neonatal rats.
OBJECTIVE
To clone and identify one novel regeneration related gene H(3) (CA854305) from the differential expression genes library we had set up before.


METHODS
Use 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.


RESULTS
Northern 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.


CONCLUSIONS
We 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.
Chu
Tong-wei
TW
Department of Orthopaedics, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. chtw@sina.com
Liu
Yu-gang
YG
Ma
Hai-han
HH
Liao
Wei-hong
WH
Wu
Ya-min
YM
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D008969:Molecular Sequence Data; D009416:Nerve Regeneration / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000235:genetics*
D018076:DNA, Complementary