摘要目的：对中国荷斯坦奶牛αs1-酪蛋白基因（CSN1S1）的可变剪接体进行鉴定，同时与NCBI上提供的参考序列进行比对分析，一方面鉴别出参考序列中确切存在的可变剪接体；另一方面在此基础上发现牛CSN1S1基因存在的新的可变剪接体种类。方法：提取泌乳期荷斯坦奶牛乳腺组织总RNA，反转录为cDNA，对牛CSN1S1基因CDS序列进行核酸分子克隆。通过构建合适有效的克隆载体，转化感受态细胞来进一步扩大培养，再根据蓝白斑筛选选取重组子，通过测序结果来分析CSN1S1基因的可变剪接体情况。结果：在本实验中一共分析鉴定出CSN1S1基因存在的10种可变剪接体。结论：本实验鉴定出中国荷斯坦奶牛CSN1S1基因的10种可变剪接体，其中4种可变剪接体与NCBI上提供的参考序列比对基本一致，其余6种预测为αs1-酪蛋白基因可能存在的新的可变剪接体。50723

该论文有图3幅，表8个，参考文献19篇。

毕业论文关键词：CSN1S1  荷斯坦奶牛  可变剪接  基因克隆

The cloning of CSN1S1 gene in cows and identification and analysis of the alternative splicing

Abstract

[Purposes] We aim to invetigate the alternative splicing of αs1-casein genes in China Holstein cows. At the same time, we want to analyze the sequences with the reference sequences which the NCBI provides by sequence alignment. On the one hand, we want to identify whether the reference sequences exactly exist alternative splicing. On the other hand, on this basis, we want to find novel alternative splicing of α-s1casein genes in Holstein cows. [Methods] We extracted total RNA of mammary tissue in lactating Holstein cows. The CDS sequences of CSN1S1 were cloned using cDNA obtained from total RNA by reverse transcription (RT) and amplified by molecular cloning. Constructing effective cloning vector, the products were transformed into Trans1-T1 competent cells, coated plates, selected monoclonal colony by blue-white selection. Then we analyzed alternative splicing of CSN1S1 by results of DNA sequencing. [Results] In this experiment, ten kinds alternative splicing of CSN1S1 have been identified. [Conclusions] In the ten kinds alternative splicing, four have been distinguished by sequence alignment with the reference sequences on NCBI, the other six are forecasted that they are new alternative splicing of CSN1S1. 

Key Words：CSN1S1; Holstein cow; Alternative splicing; Gene cloning

目录

摘要 I

Abstract II

目录 III

图清单 IV

表清单 IV

引言 1

1 材料与方法 2

1.1供试材料 2

1.2方法 3

2 结果与分析 6

2.1奶牛CSN1S1基因编码区序列的PCR扩增 6

2.2鉴定重组子 7

2.3 CSN1S1基因的测序结果 8

3 讨论 10

参考文献 12

致谢 14

图清单

图序号 图名称 页码

图2-1 奶牛CSN1S1基因编码区序列的PCR扩增结果 7

图2-2 重组质粒pMD19-T的菌液PCR鉴定结果