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人载脂蛋白CII cDNA克隆的分离、测序及其用于分离载脂蛋白CII基因并将其定位于人第19号染色体上。

Isolation and sequence of a human apolipoprotein CII cDNA clone and its use to isolate and map to human chromosome 19 the gene for apolipoprotein CII.

作者信息

Jackson C L, Bruns G A, Breslow J L

出版信息

Proc Natl Acad Sci U S A. 1984 May;81(10):2945-9. doi: 10.1073/pnas.81.10.2945.

Abstract

cDNA clones encoding human apolipoprotein CII (apo CII) were identified by screening an adult human liver cDNA library with a mixed oligonucleotide probe corresponding to all possible codons for apo CII amino acid 6-10. One clone with an approximately equal to 500-base-pair (bp) insert, designated pCII -711, was selected for DNA sequence analysis. This clone contained a DNA sequence that corresponded with the previously reported amino acid sequence of apo CII with only minor differences. The DNA sequence specified a polypeptide of 79 amino acids, compared to the 78 amino acids previously reported. The pCII -711 clone contains a 36-bp DNA sequence upstream from that specifying the NH2-terminal threonine which, when read in frame, specifies the amino acid sequence Leu-Val-Leu-Leu-Val-Leu-Gly-Phe-Glu-Val-Gln-Gly and may be part of an apo CII signal peptide. The pCII -711 clone also contains a 144-bp region that corresponds to the 3' untranslated region of apo CII mRNA as well as a portion of the poly(A) tail. Clone pCII -711 was used to isolate and characterize by restriction endonuclease digestion the gene for apo CII from a human genomic library. In addition, through Southern blot analysis of DNA from human-rodent somatic cell hybrids, clone pCII -711 also was used to provisionally map the gene for apo CII to human chromosome 19.

摘要

通过用与载脂蛋白CII(apo CII)氨基酸6 - 10的所有可能密码子对应的混合寡核苷酸探针筛选成人肝脏cDNA文库,鉴定出编码人载脂蛋白CII的cDNA克隆。选择一个插入片段约为500碱基对(bp)的克隆,命名为pCII - 711,用于DNA序列分析。该克隆包含一个与先前报道的apo CII氨基酸序列相对应的DNA序列,只有微小差异。该DNA序列指定了一个79个氨基酸的多肽,而先前报道的是78个氨基酸。pCII - 711克隆在指定氨基末端苏氨酸的序列上游含有一个36 bp的DNA序列,当按读码框读取时,指定氨基酸序列亮氨酸 - 缬氨酸 - 亮氨酸 - 亮氨酸 - 缬氨酸 - 亮氨酸 - 甘氨酸 - 苯丙氨酸 - 谷氨酸 - 缬氨酸 - 谷氨酰胺 - 甘氨酸,可能是apo CII信号肽的一部分。pCII - 711克隆还包含一个144 bp的区域,对应于apo CII mRNA的3'非翻译区以及部分聚腺苷酸尾。克隆pCII - 711用于从人基因组文库中通过限制性内切酶消化分离和鉴定apo CII基因。此外,通过对人 - 啮齿动物体细胞杂种的DNA进行Southern印迹分析,克隆pCII - 711还用于将apo CII基因初步定位到人类染色体19上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b80/345197/ee95d026f385/pnas00611-0010-a.jpg

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