香蕉ACC合成酶含3'末端的cDNA克隆

采用 ３＇ＲＡＣＥ方法对香蕉（Ｍｕｓａ ＡＡＡ Ｃａｖｅｎｄｉｓｈ Ｓｕｂｇｒｏｕｐ）ＡＣＣ合成酸含 ３＇末端的 ｃＤＮＡ进行扩增，扩增产物被克隆到ｐＣＲ■２．１载体上，转化大肠杆菌ＤＨ５ α，筛选到重组质粒（ｐＡＣＳ２），并对插入片段进行序列测定。结果表明，３＇ＲＡＣＥ产物长 １６８０ ｂｐ，其中一个 开放读框内的核苷酸序列编码４４４个氨基酸，氨基酸序列包括了ＡＣＣ合成酶中所应具有的７个高度保守区。同时该产物还有长２６９ ｂｐ的３＇末端，并具有完整的 Ｐｏｌｙ（Ａ）尾（２４ｍｅｒ）。

Cloning of a cDNA with Intact 3' end Encoding ACC Synthasein Banana (Musa in Cavendish Subgroup)

A fragment of cDNA with intact 3' end which encodes ACC synthase in banana (Musa AAA Cavendish Subgroup) was obtained through rapid amplification of cDNA 3' end (3RACE).The product of PCR was cloned in the vector pCR 2.1 and the recombinant plasmids was transformed to E.coli DH5a. The recombinant plasmid - pACS2 contains the insertion of the product ofPCR. The sequence of the PCR product showed it was 1. 680 bp long and encoded 444 amino acidswhose sequence included all six highly conserved regions of ACC synthase. The PCR product alsohad 3' end of 269 bp long with intact polyA tail (24 mer).