Germline complexity, restriction fragment length polymorphism, and coding region sequences of the human VH7 gene family identified with family-specific FR3 segment oligonucleotides.
Germline complexity, restriction fragment length polymorphism, and coding region sequences of the human VH7 gene family identified with family-specific FR3 segment oligonucleotides.
复制标题
使用家族特异性 FR3 片段寡核苷酸鉴定的人 VH7 基因家族的种系复杂性、限制性片段长度多态性和编码区序列。
DOI：
10.1016/0161-5890(94)90145-7
复制
发表时间：
1994
影响因子：
3.6
通讯作者：
Stewart,AK
中科院分区：
文献类型：
--
作者：
Rubinstein,DB;Schwartz,RS;Guillaume,T;Leblanc,P;Stewart,AK
我们使用最具家族特异性的基因片段，即框架 3 (FR3) 的 5' 端来研究最近描述的人类 VH7 基因家族的种系复杂性和编码区序列。由于VH7和VH1家族成员之间的5'序列同源性很高，全长编码区探针无法区分这两个群体。用 VH7 编码区探针与 EcoRI 消化的基因组 DNA 进行杂交，显示出 12 个片段。许多这些杂交片段也用全长 VH1 编码序列探针进行了鉴定。然而，用包含 FR3 片段 5' 端的 VH7 家族特异性寡核苷酸检查相同的 DNA 样品，大大降低了杂交复杂性，仅产生 4 个片段，其中包括一个分子量为 7.4 kb 的多态性条带。 VH7 特异性 FR3 寡核苷酸也用于在中等严格条件下从来自 6 个不相关个体的 PCR 扩增基因组 DNA 文库中分离 VH7 克隆。所有分离的克隆都含有 VH7 家族的成员。获得了六个序列。在所有个体中都可见到的基因 7A.4 与之前描述的种系 V1-4.1b 基因相同，除了 253 和 254 核苷酸处的 G-C 替换之外。还鉴定了五个不同的假基因。终止密码子限于框架2和3。先前描述的来自脐带血、正常成人和两种类风湿因子的表达的VH7基因与基因7A.4具有> 96%的同源性。
We have used the most family-specific gene segment, the 5'-end of framework 3 (FR3) to study the germline complexity and coding region sequences of the recently described human VH7 gene family. Because of the high degree of 5' sequence homology between members of the VH7 and VH1 families, full-length coding region probes are unable to distinguish between the two groups. Hybridization with a VH7 coding region probe toEcoRI digested genomic DNA revealed 12 fragments. Many of these hybridizing fragments were also identified with a full length VH1 coding sequence probe. However, examination of the same DNA samples with a VH7 family specific oligonucleotide, encompassing the 5'-end of the FR3 segment, greatly reduced hybridization complexity yielding only four fragments which included one polymorphic band of molecular weight 7.4 kb. The VH7 specific FR3 oligonucleotide was also used under conditions of moderate stringency to isolate VH7 clones from PCR-amplified genomic DNA libraries derived from six unrelated individuals. All clones isolated contained members of the VH7 family. Six sequences were obtained. Gene 7A.4, seen in all individuals, is identical to the previously described germline V1–4.1bgene other than G-C substitutions at nucleotides 253 and 254. Five distinct pseudogenes were also identified. Stop codons were confined to frameworks 2 and 3. Previously described expressed VH7 genes from cord blood, normal adults and two rheumatoid factors are > 96% homologous to gene 7A.4.
登录
查看更多内容
无需体外刺激即可从人外周血淋巴细胞构建代表性免疫球蛋白可变区 cDNA 文库。
影响因子：
2.2
作者：
Huang,C;Stollar,BD
通讯作者：
Stollar,BD
人类 Ig VH4 基因家族的多态性。
DOI：
--
发表时间：
1991
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
vanDijk,KW;Sasso,EH;Milner,EC
通讯作者：
Milner,EC
影响因子：
5
作者：
J. Berman;F. Alt
通讯作者：
F. Alt
DOI：
10.1073/pnas.87.16.6146
发表时间：
1990-08
影响因子：
11.1
作者：
H. Schroeder;Jin Yi Wang
通讯作者：
H. Schroeder;Jin Yi Wang
影响因子：
15.3
作者：
M. Walter;H. Dosch;D. Cox
通讯作者：
D. Cox