Isolation, characterization, and mapping of a human acid beta-galactosidase cDNA.
Isolation, characterization, and mapping of a human acid beta-galactosidase cDNA.
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人酸性 β-半乳糖苷酶 cDNA 的分离、表征和作图。
DOI：
10.1089/dna.1990.9.119
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发表时间：
1990
影响因子：
3.1
通讯作者：
O'Brien,JS
中科院分区：
文献类型：
--
作者：
Yamamoto,Y;Hake,CA;Martin,BM;Kretz,KA;Ahern-Rindell,AJ;Naylor,SL;Mudd,M;O'Brien,JS
根据纯化的人β-半乳糖苷酶的溴化氰片段和胰蛋白酶片段的氨基酸序列数据，用简并寡核苷酸探针混合物筛选λ gt 11人睾丸cDNA文库。筛选2 × 106个噬菌斑后，筛选出6个阳性克隆。测定了这6个克隆的序列，发现它们来源于两种不同的cDNA。这些cDNA中最长的序列与Oshimaet al.（1988年）的报告。它编码一个76 kD的蛋白质和从纯化的酶产生的所有11种肽。第二个克隆在编码区的中心部分短393 bp。通过北方印迹分析揭示了在淋巴母细胞和睾丸组织中存在2.45 kb的单个mRNA种类。从氨基酸序列数据推断，β-半乳糖苷酶前体形式的蛋白水解加工必须通过在多肽的羧基末端部分（可能在独特亲水性序列处的氨基酸530附近）中切割而发生。利用从cDNA的3′端产生的探针，我们将编码人β-半乳糖苷酶的基因定位在染色体3 p21 - 3 pter上。
A λgt11 human testicular cDNA library was screened with degenerate oligonucleotide probe mixtures based on amino acid sequence data generated from cyanogen bromide fragments and tryptic fragments of purified human β-galactosidase. Six positive clones were identified after screening 2 × 106plaques. The sequences of these six clones were determined and found to be derived from two different cDNAs. The sequence of the longest of these cDNAs is nearly identical to that recently determined by Oshimaet al.(1988). It codes for a 76-kD protein and all 11 peptides that were generated from the purified enzyme. The second clone is shorter by 393 bp in the central portion of the coding region. Analysis by Northern blotting revealed the presence of a single mRNA species of 2.45 kb in lymphoblasts and testicular tissue. It is deduced from the amino acid sequence data that proteolytic processing of the precursor form of β-galactosidase must occur by cleavage in the carboxy-terminal portion of the polypeptide perhaps around amino acid 530 at a uniquely hydrophilic sequence. Using a probe generated from the 3′ region of the cDNA, we have mapped the locus coding for human β-galactosidase to chromosome 3p21-3pter.