Cloning of Human Myelin
Protein Zero-like Genes by Bioinformatics Strategy
TANG Dong-Sheng*, YU Kuan-Ping, TANG Xi-Xiang,
ZHANG Hua-Li, PAN Qian, DAI He-Ping, XIA Jia-Hui
( National Laboratory of Medical Genetics of China, Hunan Medical
University, Changsha 410078, China )
Abstact To clone
novel myelin protein related genes, two human ESTs, which shared significant
similarity with the human myelin protein zero gene, were found by the
comparison of homologue between the cDNA coding region sequences of MPZ
gene and the EST database of NCBI. An 801 bp EST contig was assembled, which
was 100% identical with a 128 kb genomic sequence, mapped to 1q24. A 435 bp
open reading frame (ORF) within the 801 bp contig was shown by computer
analysis. Two primers，designed according to the sequence of the
contig, were coupled with the primers(λgt10-5 and gt10-5) on the sequences
flanking cloning site of the cDNA library vector to amplify the cDNA library
sequences by nested PCR. New primers, designed based on novel cDNA sequences,
were used for the PCR amplification with λgt10-5 and gt10-5 in the same way as
above. Finally, the human myelin protein zero like gene isoform I and II (MPZL1a,
MPZL1b； GenBank： AF095727, AF092424) were
cloned. Comparison of gene and protein structures between MPZL1 and MPZ
revealed that MPZL1 is the second member of MPZ family.
Mutation analysis of MPZL1 gene was performed in 24
Charcot-Marie-Tooth disease (CMT) families and 26 nonsyndrome deafness
families, but no mutation was found.
Key words myelin protein zero-like gene； in silicon cloning； gene cloning； Charcot-Marie-Tooth disease
*Corresponding author： Tel, 86-731-4472093；Fax, 86-731-4478152； e-mail, [email protected]