TOTAL SYNTHESIS OF A GENE FOR BOVINE RHODOPSIN

被引：191

作者：

FERRETTI, L ^{[1
]}

KARNIK, SS ^{[1
]}

KHORANA, HG ^{[1
]}

NASSAL, M ^{[1
]}

OPRIAN, DD ^{[1
]}

机构：

[1] MIT, DEPT CHEM, CAMBRIDGE, MA 02139 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1986年 / 83卷 / 03期

关键词：

D O I：

10.1073/pnas.83.3.599

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

To carry out systematic structure-function studies of bovine rhodopsin by specific amino acid replacements, we have accomplished the total synthesis of its gene, which is 1057 base pairs long. The synthetic gene contains 28 unique restriction sites that are on the average 60 base pairs apart. Replacement of specific restriction fragments by synthetic counterparts containing the desired nucleotide changes permits specific mutagenesis in all parts of the gene. The synthesis of the gene involved enzymatic joining of a total of 72 synthetic oligonucleotides, 15-40 nucleotides long, to form DNA duplexes. The total gene was assembled from three synthetic fragments that were cloned. All synthetic oligonucleotides were characterized by 5''-end analysis, and the accuracy of the joining reactions was confirmed by sequencing the three fragments as well as the complete gene.

引用

页码：599 / 603

页数：5

共 18 条

[1] HINDERED DIALKYLAMINO NUCLEOSIDE PHOSPHITE REAGENTS IN THE SYNTHESIS OF 2 DNA 51-MERS [J].