激光生物学报2012，Vol.21Issue(6)：517-522,509,7.DOI:10.3969/j.issn.1007-7146.2012.06.008
神经生长因子在雄鸡睾丸组织中的定量表达
Quantitative Expression of Nerve Growth Factor in the Testis of Rooster
李丽芳 1张映 2郝玉娟 2李桂兰2
作者信息
- 1. 山西农业大学信息学院,山西,太谷,030800
- 2. 山西农业大学动物科技学院,山西,太谷,030800
- 折叠
摘要
Abstract
The objective of this study was to determine and discuss the expression of nerve growth factor ( NGF ) in the testis of rooster. We took out the testes of the roosters at 20 w、24 w、28 w、35 w respectively, and froze them in liquid nitrogen , then one part of testes was used to extract total RNA with Trizol reagent. By using Primer 5. 0, we designed and synthesized specific primers according to the gallus NGF gene sequence published in GenBank. After two-step RT-PCR amplification, we performed relative quantitative analysis by Real-time PCR using β-actin as the control gene. Other testes were used to extract the total protein with RIPA tissue lysis solution for western blot. The results showed that the target gene fragment was 219 bp in size which is in line with the expected fragment size. Through homology comparison,the obtained gene sequence shared 99% identity with the NGF gene sequence of chicken in Genbank. The NGF in various week-ages roosters expressed differently at mRNA level. The NGF mRNA expression level of the roosters at sexual maturity( 20 w-28 w )increased with ages,reached the highest point at 28 w,and declined in adulthood (35 w ). The results of western blot showed that,the expression quantities of NGF increased with ages. Before 20 w,the expression levels were low, then increase apparently( 20 w-28 w ), and reached the highest point at 35 w. These results indicated that NGF mRNA expression level changed regularly with the generation and development of the rooster testis, and NGF play an important accommodation role in the development of testicular tissue and spermatogenesis. It will provide some basis for the subsequent study on the effects of NGF in the reproductive system of poultry.关键词
神经生长因子/睾丸/雄鸡/发育过程/实时荧光定量/PCRKey words
nerve growth factor/ testis/ rooster/ developmental process/ real-time PCR分类
生物科学引用本文复制引用
李丽芳,张映,郝玉娟,李桂兰..神经生长因子在雄鸡睾丸组织中的定量表达[J].激光生物学报,2012,21(6):517-522,509,7.
