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绵羊免疫球蛋白重链和轻链cDNA的分离与测序

Isolation and sequence of sheep Ig H and L chain cDNA.

作者信息

Foley R C, Beh K J

机构信息

Commonwealth Scientific and Industrial Research Organization, Division of Animal Health, Sydney, Australia.

出版信息

J Immunol. 1989 Jan 15;142(2):708-11.

PMID:2492052
Abstract

Sheep lymphocyte poly(A+) RNA was used as a template for the enzymatic synthesis of cDNA before cloning into the expression vector lambda gt11. Screening of the cDNA library with mAb probes resulted in the isolation of two recombinant phages containing Ig coding sequences of 704 bp and 925 bp. These were inserted into the EcoRI site of pUC18 and named pSLC (sheep Ig L chain) and pSHC (sheep Ig H chain). The insert in pSLC revealed sequence homology by using GenBank to lambda L chain and pSHC revealed sequence homology to IgG sequences from various species. The L chain cDNA contained the full translation sequence and 5' and 3' nontranslating region while the H chain cDNA coded for the secreted form of IgG1 and lacked sequences upstream from the C region. The derived amino acid sequences showed significant homology with various Ig sequences already described for human, mouse, rabbit, pig, and chicken but the degree of homology showed no consistency with established phylogenetic relationships.

摘要

在将绵羊淋巴细胞多聚腺苷酸(poly(A+))RNA克隆到表达载体λgt11之前,用作酶促合成cDNA的模板。用单克隆抗体(mAb)探针筛选cDNA文库,得到两个含有704bp和925bp Ig编码序列的重组噬菌体。将它们插入pUC18的EcoRI位点,分别命名为pSLC(绵羊Ig轻链)和pSHC(绵羊Ig重链)。通过使用GenBank,pSLC中的插入片段显示与λ轻链具有序列同源性,pSHC显示与来自不同物种的IgG序列具有序列同源性。轻链cDNA包含完整的翻译序列以及5'和3'非翻译区,而重链cDNA编码IgG1的分泌形式,并且缺少C区上游的序列。推导的氨基酸序列与已经描述的人、小鼠、兔、猪和鸡的各种Ig序列具有显著同源性,但同源程度与已建立的系统发育关系不一致。

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