[Screening of tissues pooled cDNA library using probes by restricted fragments of BAC positive clones of ovine MHC].
Under the premise what we have known bacterial artificial chromosome(BAC)clone sequence information and gene annotation predicted in the Chinese Merino sheep major histocompatibility complex (MHC) region, the digested fragments from 6 BAC clones that were located in the MHC region of the Chinese Merino sheep genome BAC library, which were used to screen the cDNA library using plaque in situ hybridization as probes. The full length of positive cDNA clones (sequences) isolated were completely sequenced, and the sequences obtained were aligned with the corresponding known sequence information and the BAC clones with gene annotation. Meanwhile, the sequence similarity was searched in NCBI Blastn database. This work aimed at verification of accuracy of the gene annotation results and initial analysis of gene (sequence) function. At last, 27 positive cDNA clones (sequences) in total were screened through two runs of hybridization. It was also found that these sequences could be positioned in the corresponding BAC clones, and 25 sequences were located in exon area of the annotated gene. It was verified that 23 sequences had the highest sequence similarity with those in the Bos taurus by searching against the NCBI Blastn database; moreover, the function of these sequences were closely relate to immunology.
College of Animal Science and Technology, Shihezi University, Shihezi 832003, China.
Xiao-Liang
XL
Yang
Da-Zhang
DZ
Bai
Wei
W
Qiu
Hui-Qin
HQ
Dong
Da-Quan
DQ
Li
Fang
F
Chen
Run-Lin
RL
Ma
T Blair
TB
Hugh
Jian-Feng
JF
Gao
D000818:Animals; D022202:Chromosomes, Artificial, Bacterial; D018076:DNA, Complementary / Q000737:chemistry; D015723:Gene Library; D008285:Major Histocompatibility Complex / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods; D017422:Sequence Analysis, DNA; D012756:Sheep / Q000235:genetics
D018076:DNA, Complementary