Wintero A K, Fredholm M, Davies W
Royal Veterinary and Agricultural University, Department of Animal Science and Animal Health, Division of Animal Genetics, Bülowsvej 13, 1870 Frederiksberg C, Denmark.
Mamm Genome. 1996 Jul;7(7):509-17. doi: 10.1007/s003359900153.
A porcine small intestine directionally cloned cDNA library was constructed in the vector lambda Zap II. Clones were hybridized with total labeled cDNA such that putative high-copy number transcripts could be differentiated from middle- and low-copy number transcripts prior to selection and characterization by DNA sequencing. More than 2000 non-hybridizing and 242 hybridizing clones were collected. In total, 839 clones were sequenced from the 3' end of the cDNA, and after inter-clone comparison, the unique clones were sequenced from the 5' end of the cDNA. The 5' data were used to query the sequence in databases and resulted in the identification of 630 different gene transcripts, of which 604 are new porcine genes. The identity of 361 transcripts could be identified from sequence comparison studies. The validity of this semi-random selection approach was verified by the identification of a large number of unique transcripts.
构建了一个定向克隆到λZap II载体中的猪小肠cDNA文库。将克隆与总标记cDNA进行杂交，以便在通过DNA测序进行选择和表征之前，能够区分假定的高拷贝数转录本与中、低拷贝数转录本。收集了2000多个非杂交克隆和242个杂交克隆。总共从cDNA的3'端对839个克隆进行了测序，在克隆间比较后，从cDNA的5'端对独特的克隆进行了测序。利用5'端数据查询数据库中的序列，鉴定出630种不同的基因转录本，其中604个是新的猪基因。通过序列比较研究可以鉴定出361个转录本的身份。通过鉴定大量独特的转录本，验证了这种半随机选择方法的有效性。
